Alkylamine derivative

ABSTRACT

A composition containing a compound represented by General Formula (I) below (see the definition in the specification for the symbols in the formula) or a salt thereof has an excellent CaSR agonistic effect and provides a pharmaceutical agent, a CaSR agonistic agent, a prophylactic or therapeutic agent for a disease that can be ameliorated through CaSR activation as well as seasonings and an agent for imparting kokumi.

TECHNICAL FIELD

The present invention relates to an alkylamine derivative or a saltthereof, and a pharmaceutical agent comprising the same. Moreparticularly, the present invention relates to a CaSR agonistic agent, aprophylactic or therapeutic agent for a disease that can be amelioratedthrough CaSR activation, a prophylactic or therapeutic agent forhyperparathyroidism, diarrhea and peptic ulcer, and seasonings and anagent for imparting kokumi, which have an alkylamine derivative or apharmaceutically acceptable salt thereof as an active ingredient.

BACKGROUND ART

The calcium receptor, also called the calcium sensing receptor (alsoreferred to as “CaSR”), was cloned from bovine thyroid in 1993 asG-protein coupled seven-transmembrane receptor (G-protein coupledreceptor; GPCR) that senses extracellular calcium (Ca2+) (Non-patentDocument 1). The calcium receptor has a function of altering theintracellular Ca2+ concentration by sensing extracellular Ca2+, therebyregulating production of hormones or the like involved in Ca2+ metabolicregulation, as typified by parathyroid hormone.

Recently, cinacalcet (CCT), a calcium receptor agonist, was found tohave an action of suppressing secretion of parathyroid hormone by actingon the calcium receptor of parathyroid to enhance Ca2+ sensitivity ofthe calcium receptor (Non-patent Document 2), and it has been marketedas a therapeutic drug for secondary hyperparathyroidism of dialysispatients (Non-patent Document 3).

In addition, the calcium receptor was also found to be expressed inkidney, brain, thyroid, bones and digestive tracts, and thus consideredto be involved in various diseases.

As compounds having a CaSR activating effect, other than glutathione,for example, gamma-glutamyl peptide derivatives (Non-patent Documents 4and 9), pyrrolidine derivatives (Patent Document 1) and the like areknown. In addition, CaSR agonists such as a gamma-glutamyl peptidederivative have been reported to be useful as a prophylactic ortherapeutic agent for diarrhea (Patent Document 2), a prophylactic ortherapeutic agent for acid secretion-related diseases such as gastriculcer, duodenal ulcer and reflux esophagitis (Patent Document 3), atherapeutic agent for diabetes or obesity (Patent Document 4), andfurther as an immunostimulator (Patent Document 5). Furthermore, PatentDocument 6 and Non-patent Document 9 describe that compounds having CaSRagonistic activity are also useful as agent for imparting kokumi.

These compounds, however, are structurally different from the alkylaminederivatives of the present invention.

Meanwhile, a gamma-glutamyl anilide derivative, among the alkylaminederivatives, is known to be used as a substrate for enzymatic activity(Non-patent Document 5 and Patent Document 7) as well as for itsapplication as an antimicrobial agent or an anti-allergic agent(Non-patent Document 6 and Patent Document 8) and its application as ananalytical reagent (Non-patent Documents 7 and 11). Moreover, anL-2-amino-3-N′-substituted ureidopropanoic acid derivative is known forits application as a synthetic intermediate of an asparagine analogemployed as an anticancer agent (Non-patent Document 8). The alkylaminederivative is known as a leukotriene A4 inhibitor for applicationagainst inflammatory diseases (Non-patent Document 10). The alkylaminederivative is also known for its application as an anticancer agent(Non-patent Document 12).

The above-mentioned compounds, however, have not been known for theirapplications as pharmaceutical agents with a CaSR agonistic effect orapplications as seasonings.

PRIOR ART DOCUMENTS

Patent Documents

-   [Patent Document 1] International Patent Application Publication    (pamphlet) No. WO2006/123725-   [Patent Document 2] International Patent Application Publication    (pamphlet) No. WO2008/139947-   [Patent Document 3] International Patent Application Publication    (pamphlet) No. WO2009/119554-   [Patent Document 4] International Patent Application Publication    (pamphlet) No. WO2009/107660-   [Patent Document 5] International Patent Application Publication    (pamphlet) No. WO2009/128523-   [Patent Document 6] International Patent Application Publication    (pamphlet) No. WO2007/055393-   [Patent Document 7] International Patent Application Publication    (pamphlet) No. WO2007/055393-   [Patent Document 8] Japanese Unexamined Patent Application No.    Heisei 06-172287

Non-Patent Documents

-   [Non-patent Document 1] Nature, 366: 575-580 (1993)-   [Non-patent Document 2] Current Opinion in Pharmacology, 2: 734-739    (2002)-   [Non-patent Document 3] Ethical drug package insert (5th ed.,    revised January 2010) for “REGPARA™ tablet 25 mg/REGPARA™ tablet 75    mg”-   [Non-patent Document 4] Journal of Biological Chemistry, 281(13),    8864-70 (2006)-   [Non-patent Document 5] Clinical Chemistry, 22, 2051 (1976)-   [Non-patent Document 6] Journal of Medicinal Chemistry, 8(3),    398-400 (1965)-   [Non-patent Document 7] Analytica Chimica Acta, 519(2), 181-187    (2004)-   [Non-patent Document 8] Journal of Medicinal Chemistry, 14(5),    465-466 (1971)-   [Non-patent Document 9] Journal of Biological Chemistry, 285 (2),    1016-22 (2010)-   [Non-patent Document 10] Bioorganic & Medicinal Chemistry, 16,    4863-4983 (2008)-   [Non-patent Document 11] Revue Roumaine de Chimie, 39(12), 1435-41    (1994)-   [Non-patent Document 12] J. Org. Chem., 23, 1257-1259 (1958)

SUMMARY OF THE INVENTION Problem to be Solved by the Invention

A pharmaceutical agent that has a superior CaSR agonistic effect andthat is highly safe is desired to be provided. In addition,high-performance seasonings that imparts kokumi is also desired to beprovided.

Means for Solving the Problem

As a result of search for a CaSR agonist, the present inventors foundthat an alkylamine derivative of the present invention has a superiorCaSR agonistic effect and was effective for various disease models,thereby accomplishing the present invention.

Thus, the present invention is as follows.

[1] A compound represented by the following Formula (I) or a saltthereof:

[wherein, R¹ and R², each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₂₋₆ alkenyl or substituted or unsubstituted C₂₋₆ alkynyl, or R¹ and R²may integrally form a substituted or unsubstituted 5- or 6-memberedhetero ring which may further include a heteroatom(s);

R³ represents a hydrogen atom, halogeno or substituted or unsubstitutedC₁₋₆ alkyl;

R⁴ and R⁵, each independently, represent a hydrogen atom, substituted orunsubstituted C₁₋₆ alkyl, substituted or unsubstituted C₂₋₆ alkenyl,substituted or unsubstituted C₂₋₆ alkynyl or halogeno;

X represents CR^(a)R^(b), an oxygen atom, NR^(c) or a sulfur atom(wherein, R^(a) and R^(b), each independently, represent a hydrogenatom, C₁₋₆ alkyl or halogeno, and R^(c) represents a hydrogen atom orC₁₋₆ alkyl);

Y represents C═O, SO, SO₂, C═S or C═NR^(d) (wherein R^(d) represents ahydrogen atom or C₁₋₆ alkyl, and R^(d) and R⁶ may integrally form asubstituted or unsubstituted 5- or 6-membered hetero ring);

R⁶ represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl or hydroxy;

G represents R⁷-substituted aryl or R⁷-substituted heteroaryl, where theR⁷-substituted aryl or the R⁷-substituted heteroaryl may further besubstituted with one or more R⁸;

R⁷ represents sulfo, carboxyl or phosphono;

R⁸ represents substituted or unsubstituted C₁₋₆ alkyl, substituted orunsubstituted C₂₋₆ alkenyl, substituted or unsubstituted C₂₋₆ alkynyl,halogeno, hydroxy, substituted or unsubstituted C₁₋₆ alkoxy, nitro,amino, mono-C₁₋₆ alkylamino, di-C₁₋₆ alkylamino, sulfo, carboxyl,phosphono, C₁₋₃ alkylcarbonylamino or mono-C₁₋₆ alkylphosphono, wherethey may be different when more than one R⁸ exist;

Q represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl, carboxyl, CONR^(e)R^(f), CONHNHR^(g), COR^(h), substitutedor unsubstituted aryl or substituted or unsubstituted heteroaryl;

R^(e) and R^(f), each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₁₋₆ alkylsulfonyl, substituted or unsubstituted arylsulfonyl,substituted or unsubstituted C₃₋₈ cycloalkyl, hydroxy or C₁₋₆ alkoxy, oralternatively, R^(e) and R^(f) may integrally form a substituted orunsubstituted 5- or 6-membered hetero ring which may further have aheteroatom(s);

R^(g) represents substituted or unsubstituted C₁₋₆ alkylcarbonyl,substituted or unsubstituted benzoyl, substituted or unsubstituted arylor substituted or unsubstituted heteroaryl; and

R^(h) represents substituted or unsubstituted C₁₋₆ alkoxy, substitutedor unsubstituted mercapto, or the following group:

(wherein Z represents a bivalent group of substituted or unsubstitutedC₁₋₆ hydrocarbon; E¹ represents substituted or unsubstituted C₁₋₆acyloxy, substituted or unsubstituted C₁₋₆ alkoxycarbonyloxy,substituted or unsubstituted amino, carboxyl, substituted orunsubstituted C₁₋₆ alkoxycarbonyl, halogeno, aryl, heteroaryl,substituted or unsubstituted C₁₋₆ alkoxy or substituted or unsubstitutedcarbamoyl; E² represents a hydrogen atom or C₁₋₆ alkyl; and Z and E¹ mayintegrally form a ring),

provided that when X is methylene or an oxygen atom, Y is C═O all ofR¹-R⁵ are hydrogen atoms and G is phenyl, Q is a group other thancarboxyl or COR^(h)].

[2] The compound according to [1] above, represented by the followingFormula (I), or a salt thereof:

[wherein, R¹ and R², each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₂₋₆ alkenyl or substituted or unsubstituted C₂₋₆ alkynyl, or R¹ and R²may integrally form a substituted or unsubstituted 5- or 6-memberedhetero ring which may further include a heteroatom(s);

R³ represents a hydrogen atom, halogeno or substituted or unsubstitutedC₁₋₆ alkyl;

R⁴ and R⁵, each independently, represent a hydrogen atom, substituted orunsubstituted C₁₋₆ alkyl, substituted or unsubstituted C₂₋₆ alkenyl,substituted or unsubstituted C₂₋₆ alkynyl or halogeno;

X represents CR^(a)R^(b), an oxygen atom, NR^(c) or a sulfur atom(wherein, R^(a) and R^(b), each independently, represent a hydrogenatom, C₁₋₆ alkyl or halogeno, and R^(c) represents a hydrogen atom orC₁₋₆ alkyl);

Y represents C═O, SO, SO₂, C═S or C═NR^(d) (wherein R^(d) represents ahydrogen atom or C₁₋₆ alkyl, and R^(d) and R⁶ may integrally form asubstituted or unsubstituted 5- or 6-membered hetero ring);

R⁶ represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl or hydroxy;

G represents R⁷-substituted aryl or R⁷-substituted heteroaryl, where theR⁷-substituted aryl or the R⁷-substituted heteroaryl may further besubstituted with one or more R⁸;

R⁷ represents sulfo, carboxyl or phosphono;

R⁸ represents substituted or unsubstituted C₁₋₆ alkyl, substituted orunsubstituted C₂₋₆ alkenyl, substituted or unsubstituted C₂₋₆ alkynyl,halogeno, hydroxy, substituted or unsubstituted C₁₋₆ alkoxy, nitro,amino, mono-C₁₋₆ alkylamino, di-C₁₋₆ alkylamino, sulfo, carboxyl,phosphono or mono-C₁₋₆ alkylphosphono, where they may be different whenmore than one R⁸ exist;

Q represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl, carboxyl, CONR^(e)R^(f), CONHNHR^(g), COR^(h), substitutedor unsubstituted aryl or substituted or unsubstituted heteroaryl;

R^(e) and R^(f), each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₁₋₆ alkylsulfonyl, substituted or unsubstituted arylsulfonyl,substituted or unsubstituted C₃₋₈ cycloalkyl, hydroxy or C₁₋₆ alkoxy, oralternatively, R^(e) and R^(f) may integrally form a substituted orunsubstituted 5- or 6-membered hetero ring which may further have aheteroatom(s);

R^(g) represents substituted or unsubstituted C₁₋₆ alkylcarbonyl,substituted or unsubstituted benzoyl, substituted or unsubstituted aryl,or substituted or unsubstituted heteroaryl; and

R^(h) represents substituted or unsubstituted C₁₋₆ alkoxy, substitutedor unsubstituted mercapto, or the following group:

(wherein Z represents a bivalent group of substituted or unsubstitutedC₁₋₆ hydrocarbon; E¹ represents substituted or unsubstituted C₁₋₆acyloxy, substituted or unsubstituted C₁₋₆ alkoxycarbonyloxy,substituted or unsubstituted amino, carboxyl, substituted orunsubstituted C₁₋₆ alkoxycarbonyl, halogeno, aryl, heteroaryl,substituted or unsubstituted C₁₋₆ alkoxy or substituted or unsubstitutedcarbamoyl; E² represents a hydrogen atom or C₁₋₆ alkyl; and Z and E¹ mayintegrally form a ring),

provided that when X is methylene or an oxygen atom, Y is C═O, all ofR¹-R⁵ are hydrogen atoms and G is phenyl, Q is a group other thancarboxyl or COR^(h)].

[3] A pharmaceutical agent comprising the compound or a pharmaceuticallyacceptable salt thereof according to [1] or [2] above as an activeingredient.

[4] A CaSR agonistic agent comprising a compound represented by thefollowing Formula (I⁰) or a pharmaceutically acceptable salt thereof asan active ingredient:

[wherein, R¹ and R², each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₂₋₆ alkenyl or substituted or unsubstituted C₂₋₆ alkynyl, or R¹ and R²may integrally form a substituted or unsubstituted 5- or 6-memberedhetero ring which may further include a heteroatom(s);

R³ represents a hydrogen atom, halogeno or substituted or unsubstitutedC₁₋₆ alkyl;

R⁴ and R⁵, each independently, represent a hydrogen atom, substituted orunsubstituted C₁₋₆ alkyl, substituted or unsubstituted C₂₋₆ alkenyl,substituted or unsubstituted C₂₋₆ alkynyl or halogeno;

X represents CR^(a)R^(b), an oxygen atom, NR^(c) or a sulfur atom(wherein, R^(a) and R^(b), each independently, represent a hydrogenatom, C₁₋₆ alkyl or halogeno, and R^(c) represents a hydrogen atom orC₁₋₆ alkyl);

Y represents C═O, SO, SO₂, C═S or C═NR^(d) (wherein R^(d) represents ahydrogen atom or C₁₋₆ alkyl, and R^(d) and R⁶ may integrally form asubstituted or unsubstituted 5- or 6-membered hetero ring);

R⁶ represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl or hydroxy;

G⁰ represents aryl that is unsubstituted or substituted with one or moreR⁷⁰ or heteroaryl that is unsubstituted or substituted with one or moreR⁷⁰, where the R⁷⁰-substituted aryl or the R⁷⁰-substituted heteroarylmay further be substituted;

R⁷⁰ represents substituted or unsubstituted C₁₋₆ alkyl, substituted orunsubstituted C₂₋₆ alkenyl, substituted or unsubstituted C₂₋₆ alkynyl,halogeno, hydroxy, substituted or unsubstituted C₁₋₆ alkoxy, nitro,amino, mono-C₁₋₆ alkylamino, di-C₁₋₆ alkylamino, sulfo, carboxyl,phosphono, C₁₋₃ alkylcarbonylamino or mono-C₁₋₆ alkylphosphono, wherethey may be different when more than one R⁷⁰ exist;

Q represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl, carboxyl, CONR^(e)R^(f), CONHNHR^(g), COR^(h), substitutedor unsubstituted aryl or substituted or unsubstituted heteroaryl;

R^(e) and R^(f), each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₁₋₆ alkylsulfonyl, substituted or unsubstituted arylsulfonyl,substituted or unsubstituted C₃₋₈ cycloalkyl, hydroxy or C₁₋₆ alkoxy, oralternatively, R^(e) and R^(f) may integrally form a substituted orunsubstituted 5- or 6-membered hetero ring which may further have aheteroatom(s);

R^(g) represents substituted or unsubstituted C₁₋₆ alkylcarbonyl,substituted or unsubstituted benzoyl, substituted or unsubstituted aryl,or substituted or unsubstituted heteroaryl; and

R^(h) represents substituted or unsubstituted C₁₋₆ alkoxy, substitutedor unsubstituted mercapto, or the following group:

(wherein Z represents a bivalent group of substituted or unsubstitutedC₁₋₆ hydrocarbon; E′ represents substituted or unsubstituted C₁₋₆acyloxy, substituted or unsubstituted C₁₋₆ alkoxycarbonyloxy,substituted or unsubstituted amino, carboxyl, substituted orunsubstituted C₁₋₆ alkoxycarbonyl, halogeno, aryl, heteroaryl,substituted or unsubstituted C₁₋₆ alkoxy or substituted or unsubstitutedcarbamoyl; E² represents a hydrogen atom or C₁₋₆ alkyl; and Z and E¹ mayintegrally form a ring),

provided that when X is methylene or an oxygen atom, Y is C═O, all ofR¹-R⁵ are hydrogen atoms, and G is phenyl, then, Q is a group other thancarboxyl or COR^(h)].

[5] The CaSR agonistic agent according to [4] above, comprising acompound represented by the following Formula (I⁰) or a pharmaceuticallyacceptable salt thereof as an active ingredient:

[wherein, R¹ and R², each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₂₋₆ alkenyl or substituted or unsubstituted C₂₋₆ alkynyl, or R¹ and R²may integrally form a substituted or unsubstituted 5- or 6-memberedhetero ring which may further include a heteroatom(s);

R³ represents a hydrogen atom, halogeno or substituted or unsubstitutedC₁₋₆ alkyl;

R⁴ and R⁵, each independently, represent a hydrogen atom, substituted orunsubstituted C₁₋₆ alkyl, substituted or unsubstituted C₂₋₆ alkenyl,substituted or unsubstituted C₂₋₆ alkynyl or halogeno;

X represents CR^(a)R^(b), an oxygen atom, NR^(c) or a sulfur atom(wherein, R^(a) and R^(b), each independently, represent a hydrogenatom, C₁₋₆ alkyl or halogeno, and R^(c) represents a hydrogen atom orC₁₋₆ alkyl);

Y represents C═O, SO, SO₂, C═S or C═NR^(d) (wherein R^(d) represents ahydrogen atom, C₁₋₆ alkyl, R^(d) and R⁶ may integrally form asubstituted or unsubstituted 5- or 6-membered hetero ring);

R⁶ represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl or hydroxy;

G⁰ represents unsubstituted or substituted aryl with one or more R⁷⁰ orunsubstituted or substituted heteroaryl with one or more R⁷⁰, where theR⁷⁰-substituted aryl or the R⁷⁰-substituted heteroaryl may further besubstituted;

R⁷⁰ represents substituted or unsubstituted C₁₋₆ alkyl, substituted orunsubstituted C₂₋₆ alkenyl, substituted or unsubstituted C₂₋₆ alkynyl,halogeno, hydroxy, substituted or unsubstituted C₁₋₆ alkoxy, nitro,amino, mono-C₁₋₆ alkylamino, di-C₁₋₆ alkylamino, sulfo, carboxyl,phosphono or mono-C₁₋₆ alkylphosphono, where they may be different whenmore than one R⁷⁰ exist;

Q represents a hydrogen atom, substituted or unsubstituted C₁₋₆ alkyl,substituted or unsubstituted C₂₋₆ alkenyl, substituted or unsubstitutedC₂₋₆ alkynyl, carboxyl, CONR^(e)R^(f), CONHNHR^(g), COR^(h), substitutedor unsubstituted aryl or substituted or unsubstituted heteroaryl;

R^(e) and R^(f), each independently, represent a hydrogen atom,substituted or unsubstituted C₁₋₆ alkyl, substituted or unsubstitutedC₁₋₆ alkylsulfonyl, substituted or unsubstituted arylsulfonyl,substituted or unsubstituted C₃₋₈ cycloalkyl, hydroxy or C₁₋₆ alkoxy, oralternatively, R^(e) and R^(f) may integrally form a substituted orunsubstituted 5- or 6-membered hetero ring which may further have aheteroatom(s);

R^(h) represents substituted or unsubstituted C₁₋₆ alkylcarbonyl,substituted or unsubstituted benzoyl, substituted or unsubstituted aryl,or substituted or unsubstituted heteroaryl; and

R^(h) represents substituted or unsubstituted C₁₋₆ alkoxy, substitutedor unsubstituted mercapto, or the following group:

(wherein Z represents a bivalent group of substituted or unsubstitutedC₁₋₆ hydrocarbon; E¹ represents substituted or unsubstituted C₁₋₆acyloxy, substituted or unsubstituted C₁₋₆ alkoxycarbonyloxy,substituted or unsubstituted amino, carboxyl, substituted orunsubstituted C₁₋₆ alkoxycarbonyl, halogeno, aryl, heteroaryl,substituted or unsubstituted C₁₋₆ alkoxy or substituted or unsubstitutedcarbamoyl; E² represents a hydrogen atom or C₁₋₆ alkyl; and Z and E¹ mayintegrally form a ring),

provided that when X is methylene or an oxygen atom, Y is C═O, all ofR¹-R⁵ are hydrogen atoms, and G is phenyl, then, Q is a group other thancarboxyl or COR^(h)].

[5-2] The CaSR agonistic agent according to either one of [4] and [5]above, which is a prophylactic or therapeutic agent forhyperparathyroidism.

[5-3] The CaSR agonistic agent according to either one of [4] and [5]above, which is a prophylactic or therapeutic agent for diarrhea.

[5-4] The CaSR agonistic agent according to either one of [4] and [5]above, which is a prophylactic or therapeutic agent for peptic ulcer.

[5-5] The CaSR agonistic agent according to either one of [4] and [5]above, which is an agent for imparting kokumi.

[6] The pharmaceutical agent according to [3] above, which is aprophylactic or therapeutic agent for a disease that is amelioratedthrough CaSR activation.

[7] The pharmaceutical agent according to [3] above, which is aprophylactic or therapeutic agent for hyperparathyroidism.

[8] The pharmaceutical agent according to [3] above, which is aprophylactic or therapeutic agent for diarrhea.

[9] The pharmaceutical agent according to [3] above, which is aprophylactic or therapeutic agent for peptic ulcer.

[10] Seasonings comprising the compound or an edible salt thereofaccording to [1] or [2] above as an active ingredient.

[11] A agent for imparting kokumi comprising the compound or an ediblesalt thereof according to either one of [1] and [2] above as an activeingredient.

Effect of the Invention

An alkylamine derivative of the present invention has a superior CaSRagonistic effect, and useful, for example, as a prophylactic ortherapeutic agent for a disease that is ameliorated through CaSRactivation, in particular, as a prophylactic or therapeutic agent forhyperparathyroidism, diarrhea or peptic ulcer, and as seasonings or anagent for imparting kokumi.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 A graph comparing the effects of Compound No. 1 and cinacalcetwith respect to serum iPTH concentration.

FIG. 2 A graph comparing the effects of Compound No. 1 and cinacalcetwith respect to serum Ca concentration.

FIG. 3 A graph showing the effects of Compounds Nos. 1 and 2 onNSAID-induced small intestine inflammation. (*P<0.05)

FIG. 4 A graph showing the effect of Compound No. 3 on NSAID-inducedsmall intestine inflammation.

FIG. 5 A graph showing the effect of Compound No. 1 with respect towater absorption action using a colon loop technique.

FIG. 6 A graph showing the effect of Compound No. 2 with respect towater absorption action using a colon loop technique.

FIG. 7 A graph showing the effect of Compound No. 3 with respect towater absorption action using a colon loop technique.

FIG. 8 A graph showing the effect of Compound No. 4 with respect towater absorption action using a colon loop technique.

EMBODIMENTS FOR CARRYING OUT THE INVENTION

Hereinafter, definitions of the groups of the compounds represented byFormulae (I) and (I⁰) will be described.

Herein, “C₁₋₆ alkyl” is a monovalent group derived by removing any onehydrogen atom from a linear- or branched-chain aliphatic hydrocarbonhaving 1-6 carbons. Specific examples include methyl, ethyl, isopropyl,butyl, n-butyl, isobutyl, sec-butyl, t-butyl, pentyl, isopentyl,2,3-dimethylpropyl and hexyl. Preferably, it is C₁₋₃ alkyl.

“C₂₋₆ alkenyl” is a monovalent group with at least one double bond (twoadjacent sp2 carbon atoms) among the linear- or branched-chain aliphatichydrocarbon groups having 1-6 carbons. Specific examples of C₂₋₆ alkenylinclude vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl(including cis and trans), 3-butenyl, pentenyl and hexenyl. Preferably,it is C₂₋₃ alkenyl.

“C₂₋₆ alkynyl” is a monovalent group with at least one triple bond (twoadjacent sp carbon atoms) among the linear- or branched-chain aliphatichydrocarbon groups having 1-6 carbons. Specific examples includeethynyl, 1-propynyl, propargyl and 3-butynyl. Preferably, it may be C₂₋₃alkynyl.

“Halogeno” refers to fluorine, chlorine, bromine, iodine atoms and thelike.

“Aryl” refers to an aromatic hydrocarbon ring group such as phenyl andnaphthyl. Preferably, it is phenyl.

“Heteroaryl” refers to a 5- to 10-membered aromatic hetero ring groupcontaining one to four heteroatoms selected from N, S and O, Specificexamples of the aromatic hetero ring include pyridine, pyridazine,pyrazine, pyrimidine, thiazole, isothiazole, oxazole, isooxazole,oxadiazole, pyrazole, imidazole, furan, thiophene and pyrrol.Preferably, it is pyridine, imidazole, thiophene, oxadiazole or indole.It is preferably a 5- to 6-membered aromatic hetero ring andparticularly pyridine or pyrimidine.

“C₁₋₆ alkoxy” refers to C₁₋₆ alkyl-O—. Specifically, examples includemethoxy, ethoxy, 1-propoxy, 2-propoxy, n-butoxy, i-butoxy, sec-butoxy,t-butoxy, 1-pentyloxy, 2-pentyloxy, 3-pentyloxy, 2-methyl-1-butyloxy,3-methyl-1-butyloxy, 2-methyl-2-butyloxy, 3-methyl-2-butyloxy,2,2-dimethyl-1-propyloxy, 1-hexyloxy, 2-hexyloxy and 3-hexyloxy.Preferably, it is C₁₋₃ alkoxy.

Examples of “C₃₋₈ cycloalkyl” include cyclopropyl, cyclopentyl,cyclohexyl and cycloheptyl. Preferably, it is C₅₋₇ cycloalkyl.

“Mono-C₁₋₆ alkylamino” is an amino group having one hydrogen atom on thenitrogen atom substituted with the above-described C₁₋₆ alkyl, andrefers to C₁₋₆ alkyl-NH—. Specific examples include methylamino andethylamino. Preferably, it is mono-C₁₋₃ alkylamino.

“Di-C₁₋₆ alkylamino” is an amino group where each of two hydrogen atomson the nitrogen atom is substituted with the above-described C₁₋₆ alkyl,and refers to (C₁₋₆ alkyl)₂N—. The C₁₋₆ alkyl groups may be identical toor different from each other. Specific examples include dimethylaminoand diethylamino. Preferably, it is di-C₁₋₃ alkylamino.

“C₁₋₃ alkylcarbonylamino” refers to a group represented by C₁₋₃alkyl-C(O)—NH—. Examples of C₁₋₃ alkylcarbonylamino include groups suchas acetylamino and propionylamino. Preferably, it is acetylamino.

“Mono-C₁₋₆ alkylphosphono” is a phosphono group where one hydrogen atomon the hydroxyl group is substituted with the above-described C₁₋₈alkyl, and refers to —PO₃H (C₁₋₆ alkyl). Specific examples includemethylphosphono and ethylphosphono. Preferably, it is mono-C₁₋₃alkylphosphono.

“C₁₋₆ alkylsulfonyl” refers to a group represented by C₁₋₆ alkyl-S(O)₂—,and specific examples include methylsulfonyl and ethylsulfonyl.Preferably, it is C₁₋₃ alkylsulfonyl.

“Arylsulfonyl” refers to a group represented by aryl-S(O)₂—, and aspecific example includes phenylsulfonyl.

“C₁₋₆ alkylcarbonyl” refers to a group represented by C₁₋₆ alkyl-C(O)—,and specific examples include methylcarbonyl and ethylcarbonyl.Preferably, it is C₁₋₃ alkylcarbonyl.

“C₁₋₆ alkoxycarbonyloxy” refers to a group represented by C₁₋₆alkyl-O—C(O)—O—, and examples include methoxycarbonyloxy andethoxycarbonyloxy. Preferably, it is C₁₋₃ alkoxycarbonyloxy.

A “bivalent group of C₁₋₆ hydrocarbon” refers to a bivalent groupderived by removing any two hydrogen atoms from a linear- orbranched-chain aliphatic hydrocarbon that has 1-6 carbons and that maycontain one to several double or triple bonds. Specific examples includemethylene, ethane-1,1-diyl, vinylene, ethynylene and propargyl.

“C₁₋₆ acyloxy” refers to a group represented by C₁₋₆ alkyl-C(O)—O—, C₃₋₆cycloalkyl-C(O)—O— or aryl-C(O)—O—. Examples of C₁₋₆ acyloxy includegroups such as acetyloxy, propionyloxy, cyclohexylcarbonyloxy andbenzoyloxy. It is preferably C₁₋₆ alkyl-C(O)—O— and more preferably C₁₋₃alkyl-C(O)—O—.

“C₁₋₆ alkoxycarbonyl” refers to a group represented by C₁₋₆alkyl-O—C(O)—, and examples include methoxycarbonyl and ethoxycarbonyl.Preferably, it is C₁₋₃ alkoxycarbonyl.

“5- or 6-membered hetero ring that may further contain a heteroatom(s)”,formed integrally with R¹ and R² or R^(e) and R^(f), refers to asaturated or unsaturated 5- or 6-membered hetero ring that may furtherhave, other than the nitrogen atom bound by R¹ and R² or R^(e) andR^(f), 1-3 heteroatoms selected from a nitrogen atom, an oxygen atom anda sulfur atom as constituent atoms of the ring

Examples of the saturated 5- or 6-membered hetero ring group includepyrrolidine-1-yl, pyrazolidine-1-yl, imidazolidine-1-yl,piperidine-1-yl, piperazine-1-yl, morpholine-4-yl andthiomorpholine-4-yl.

Examples of the unsaturated 5- or 6-membered hetero ring group includepyrrol-1-yl, 2-pyrroline-1-yl, 3-pyrroline-1-yl, pyrazole-1-yl,imidazole-1-yl, 2-pyrazoline-1-yl, 3-pyrazoline-1-yl,2-imidazoline-1-yl, 1,2,3-triazole-1-yl, 1,2,4-triazole-1-yl,tetrazole-1-yl, 1,4-oxazine-4-yl and 1,4-thiazine-1-yl.

A “5- or 6-membered hetero ring” formed integrally with R^(d) and R⁶refers to a saturated or unsaturated 5- or 6-membered hetero ring whichincludes N═C—N bound by R^(d) and R⁶ as a part of the ring, and mayfurther contain 1-3 heteroatoms selected from a nitrogen atom, an oxygenatom and a sulfur atom as constituent atoms of the ring. Specificexamples include 2-imidazolidine, imidazole, triazole, tetrazole,1,4,5,6-tetrahydropyrimidine, 1,4-dihydropyrimidine,1,6-dihydropyrimidine and 2H-1,2,4-oxadiazine.

The phrase “which may further substituted” used for G° means thatsubstitution, for example, with a substituent such as cyano, substitutedor unsubstituted mercapto, C₁₋₆ alkyl-C(O)—, C₁₋₆ alkyl-C(O)—O—, C₃₋₆cycloalkyl, C₁₋₆ alkyl-O—C(O)—C₁₋₆ alkylene-O—, substituted orunsubstituted aryl, substituted or unsubstituted aryl-O—, substituted orunsubstituted aryl-C(O)—, —O—C₁₋₆ alkylene-O—, substituted orunsubstituted aryl-C₁₋₆ alkenyl- may take place at a position which mayhave a substituent(s) other than R⁷⁰.

Here, C₁₋₆ alkylene refers to a bivalent group of the above-describedC₁₋₆ alkyl.

When R^(h) in CO—R^(h) of Q is represented by Formula (IIa), the “ring”integrally formed with Z and E¹ refers to a saturated or unsaturated 5-or 6-membered ring which contains Z-E¹ as a part of the ring, which mayfurther have 1-3 heteroatoms selected from a nitrogen atom, an oxygenatom and a sulfur atom as constituent atoms of the ring, and which mayfurther condense with a benzene ring. Preferably, it is a saturated orunsaturated 5- or 6-membered ring which may have 1-3 oxygen atoms asconstituent atoms of the ring.

Specific examples of R^(h) where the ring is formed integrally with Zand E¹ include the following groups.

Here, the group represented by the above-described CO—R^(h) may alsorepresent a carboxyl group which has been subjected to prodrugmodification so that it is converted into a carboxyl group in vivo asdescribed, for example, in Prog. Med. 5: 2157-2161 (1985), “MolecularDesign”, Iyakuhin No Kaihatsu (Development of Pharmaceutical Product)Vol. 7, p. 163-198 (Hirokawa Shoten Co., 1990), or Saisin Soyaku Kagaku(The Practice of Medicinal Chemistry) Vol. 2, p. 271-298 (Technomics,1999).

Examples of substituents for substituted or unsubstituted C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ alkoxy, C₁₋₆ alkylsulfonyl, C₁₋₆alkylcarbonyl, mercapto, carbamoyl, amino, C₁₋₆ alkoxycarbonyl, thebivalent group of C₁₋₆ hydrocarbon, C₁₋₆ alkoxycarbonyloxy or C₁₋₆acyloxy include a halogen atom (for example, a chlorine atom, a bromineatom and a fluorine atom), hydroxy, cyano, C₁₋₆ alkoxy (for example,methoxy), C₁₋₆ halogenoalkyl (for example, trifluoromethyl),unsubstituted or mono- or di-substituted carbamoyl with C₁₋₆ alkyl,unsubstituted or substituted aryl with 1 to 3 halogeno, C₁₋₆ alkyl, C₁₋₆alkoxy or the like, and unsubstituted or substituted heteroaryl with 1to 3 halogeno, C₁₋₆ alkyl, C₁₋₆ alkoxy or the like. Preferably, it iscyano, unsubstituted or mono- or di-substituted carbamoyl with C₁₋₆alkyl, unsubstituted or substituted aryl with 1 to 3 halogeno, C₁₋₆alkyl, C₁₋₆ alkoxy or the like, or unsubstituted or substitutedheteroaryl with 1 to 3 halogeno, C₁₋₆ alkyl, C₁₋₆ alkoxy or the like.The above-mentioned groups may be 1- to 3-substituted with anysubstituent selected from these substituents at positions which may havea substituent(s). When there are multiple substitutions, thesubstituents may be different from each other.

Examples of the substituents for substituted or unsubstituted 5- or6-membered hetero ring, substituted or unsubstituted aryl, heteroaryl orarylsulfonyl or benzoyl include a halogen atom (for example, a chlorineatom, a bromine atom and a fluorine atom), hydroxy, cyano, C₁₋₆ alkyl(for example, methyl or benzyl) that may be substituted with aryl (forexample, phenyl), C₁₋₆ alkoxy (for example, methoxy), C₁₋₆ halogenoalkyl(for example, trifluoromethyl), carbamoyl that is optionally mono- ordi-substituted with C₁₋₆ alkyl, and heteroaryl that may be substitutedwith halogen or C₁₋₆ alkyl. Preferable examples include C₁₋₆ alkyl thatmay be substituted with aryl (for example, phenyl) and heteroaryl thatmay be substituted with halogen or C₁₋₆ alkyl. The above-describedgroups may be 1- to 3-substituted with any substituent selected fromthese substituents at positions which may have a substituent(s). Whenthere are multiple substitutions, the substituents may be different fromeach other

Other than the substituents enumerated above as substituents for thesubstituted or unsubstituted 5- or 6-membered hetero ring, thesubstituent for substituted or unsubstituted C₃₋₆ cycloalkyl alsoinclude oxo.

R¹ and R² are preferably a hydrogen atom or C₁₋₆ alkyl, and morepreferably a hydrogen atom.

R³ is preferably a hydrogen atom, halogeno or C₁₋₆ alkyl, and morepreferably a hydrogen atom.

R⁴ and R⁵ are preferably, a hydrogen atom, C₁₋₆ alkyl or halogeno, andmore preferably a hydrogen atom.

X is preferably CH₂, an oxygen atom, NH or a sulfur atom, and morepreferably NH or a sulfur atom. Particularly preferably, it is NH.

Y is preferably C═O, SO, SO₂ or C═S, and more preferably, C═O or C═S.

R⁶ is preferably a hydrogen atom, hydroxy or C₁₋₆ alkyl, and morepreferably a hydrogen atom.

G is preferably R⁷-substituted aryl or R⁷-substituted heteroaryl, andmay further be substituted with one to three R⁸. More preferably, it isR⁷-substituted phenyl or R⁷-substituted pyridyl, and may further besubstituted with one to two R⁸.

In the case where G is R⁷-substituted phenyl where R⁷ is sulfo, thesubstitution position of R⁷ is preferably the 3-position on the phenylgroup (wherein the carbon of the phenyl group which binds to thenitrogen in General Formula (I) is the 1-position). More specifically, Gis preferably 5-chloro-2-hydroxy-3-sulfophenyl,3-chloro-2-methyl-5-sulfophenyl or 3-chloro-4-methyl-5-sulfophenyl.

G⁰ preferably represents unsubstituted or substituted aryl with one tofive R⁷⁰ or unsubstituted or substituted heteroaryl with one to fiveR⁷⁰, and more preferably unsubstituted or substituted aryl with one tothree R⁷⁰ or unsubstituted or substituted heteroaryl with one to threeR⁷⁰, and most preferably unsubstituted or substituted phenyl with one tothree R⁷⁰ or unsubstituted or substituted pyridyl with one to three R⁷⁰.

In the case where G⁰ is R⁷⁰-substituted phenyl and one of R⁷⁰ is sulfo,the substitution position of R⁷⁰ is preferably the 3-position on thephenyl group (wherein the carbon in the phenyl group which binds withnitrogen in General Formula (I⁰) is the 1-position).

R⁷ is preferably, sulfo or carboxyl, and more preferably sulfo.

R⁷⁰ is preferably C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, halogeno,hydroxy, C₁₋₆ alkoxy, nitro, amino, mono-C₁₋₆ alkylamino, di-C₁₋₆alkylamino or mono-C₁₋₆ alkylphosphono, more preferably C₁₋₆ alkyl,halogeno, hydroxy, C₁₋₆ alkoxy, nitro, sulfo, carboxyl or phosphono, andmost preferably, C₁₋₆ alkyl, halogeno, hydroxy or sulfo.

R⁸ is preferably C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, halogeno,hydroxy, C₁₋₆ alkoxy, nitro, amino, mono-C₁₋₆ alkylamino, di-C₁₋₆alkylamino, sulfo, carboxyl, phosphono or mono-C₁₋₆ alkylphosphono, morepreferably C₁₋₆ alkyl, halogeno, hydroxy, nitro or sulfo, and mostpreferably C₁₋₆ alkyl, halogeno or hydroxy.

Q is preferably a hydrogen atom, substituted or unsubstituted C₁₋₆alkyl, carboxyl, CONR^(e)R^(f), CONHNHR^(g), COR^(h), aryl orsubstituted heteroaryl, more preferably unsubstituted or substitutedC₁₋₆ alkyl with cyano, carbamoyl or aryl, carboxyl, CONR^(e)R^(f),CONHNHR^(g), COR^(h), aryl or substituted heteroaryl with C₁₋₆ alkyl,aryl-substituted C₁₋₆ alkyl, C₁₋₆ alkyl, halogeno-substituted aryl orheteroaryl, and most preferably carboxyl.

In the case where Q is a carboxyl group, R³ is a hydrogen atom and X isCR^(a)R^(b), an oxygen atom or NR^(c), then, the steric structure of thecarbon atoms bound with Q and R³ preferably takes S-configuration.Moreover, in the case where Q is a carboxyl group, R³ is a hydrogen atomand X is a sulfur atom, then, the steric structure of the carbon atomsbound with Q and R³ preferably takes R-configuration.

R^(e) and R^(f) are preferably a hydrogen atom, substituted orunsubstituted C₁₋₆ alkyl, C₁₋₆ alkylsulfonyl, arylsulfonyl, C₃₋₆cycloalkyl, hydroxy or C₁₋₆ alkoxy, more preferably a hydrogen atom,unsubstituted or substituted C₁₋₆ alkyl (with heteroaryl, aryl, halogenor C₁₋₆ alkoxy-substituted aryl), C₁₋₆ alkylsulfonyl, arylsulfonyl,hydroxy or C₁₋₆ alkoxy, and most preferably a hydrogen atom,unsubstituted or substituted C₁₋₆ alkyl (with heteroaryl or aryl), C₁₋₆alkylsulfonyl or hydroxy.

R^(g) is preferably substituted or unsubstituted C₁₋₆ alkylcarbonyl andmore preferably unsubstituted or substituted C₁₋₆ alkylcarbonyl (witharyl).

R^(h) is preferably C₁₋₆ alkoxy, mercapto, or the following group:

(wherein, Z represents a bivalent group of C₁₋₆ hydrocarbon, E¹represents C₁₋₆ acyloxy, C₁₋₆ alkoxycarbonyloxy, amino, carboxyl, C₁₋₆alkoxycarbonyl, halogeno, aryl, heteroaryl, C₁₋₆ alkoxy or carbamoyl, E²represents a hydrogen atom or C₁₋₆ alkyl, Z and E¹ may integrally formthe following group:

Among the compounds according to [1] above, the compound (I) of thepresent invention is preferably a compound represented as follows, or asalt thereof:

R¹ and R², each independently, represent a hydrogen atom, substituted orunsubstituted C₁₋₆ alkyl, substituted or unsubstituted C₂₋₆ alkenyl orsubstituted or unsubstituted C₂₋₆ alkynyl;

R⁴ and R⁵, each independently, represent a hydrogen atom;

X represents CH₂, an oxygen atom, NH or a sulfur atom;

Y represents C═O, SO, SO₂ or C═S;

G represents R⁷-substituted aryl or R⁷-substituted heteroaryl, where theR⁷-substituted aryl or the R⁷-substituted heteroaryl may further besubstituted with one to five R⁸;

R⁸ represents substituted or unsubstituted C₁₋₆ alkyl, substituted orunsubstituted C₂₋₆ alkenyl, or substituted or unsubstituted C₂₋₆alkynyl, halogeno, hydroxy, substituted or unsubstituted C₁₋₆ alkoxy,amino, mono-C₁₋₆ alkylamino, di-C₁₋₆ alkylamino, sulfo, carboxyl,phosphono or mono-C₁₋₆ alkylphosphono, where they may be different whenmore than one R⁸ exist; and

Q is substituted or unsubstituted C₁₋₆ alkyl, substituted orunsubstituted C₂₋₆ alkenyl, substituted or unsubstituted C₂₋₆ alkynyl,carboxyl, CONR^(e)R^(f), CONHNHR^(g), substituted or unsubstituted arylor substituted or unsubstituted heteroaryl.

Furthermore, among the compounds according to [1] above, the compound(I) of the present invention is more preferably a compound representedas follows, or a salt thereof:

R¹, R², R³, R⁴, R⁵ and R⁶ are hydrogen atoms;

X is NH or a sulfur atom;

Y is C═O or C═S;

Q is carboxyl or C₁₋₆ alkoxycarbonyl;

G represents R⁷-substituted aryl or R⁷-substituted heteroaryl, where theR⁷-substituted aryl or the R⁷-substituted heteroaryl may further besubstituted with one to five R⁸;

R⁷ is sulfo;

R⁸ represents substituted or unsubstituted C₁₋₆ alkyl, substituted orunsubstituted C₂₋₆ alkenyl, or substituted or unsubstituted C₂₋₆alkynyl, halogeno, hydroxy, substituted or unsubstituted C₁₋₆ alkoxy,amino, mono-C₁₋₆ alkylamino, di-C₁₋₆ alkylamino, sulfo, carboxyl,phosphono or mono-C₁₋₆ alkylphosphono, where they may be different whenmore than one R⁸ exist.

Meanwhile, a compound represented by General Formula (I) wherein whenR¹, R², R³, R⁴, R⁵ and R⁶ are hydrogen atoms, X is methylene, Y is C═Oand G is carboxyl-substituted pyridyl, Q is not methoxycarbonyl ispreferable as a compound of the present invention.

A particularly preferably compound is a compound selected from below ora salt thereof:

-   (2R)-2-amino-3-{[(3-sulfophenyl)carbamoyl]sulfanyl}propanoic acid;-   (2S)-2-amino-3-{[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]amino}propanoic    acid;-   (2S)-2-amino-3-{[(3-chloro-4-methyl-5-sulfophenyl)carbamoyl]amino}propanoic    acid;-   (2S)-2-amino-3-{[(3-chloro-2-methyl-5-sulfophenyl)carbamoyl]amino}propanoic    acid;-   (2S)-2-amino-3-{[(3-sulfophenyl)carbamothioyl]amino}propanoic acid;-   (2S)-2-amino-3-{[(3-chloro-2-methyl-5-sulfophenyl)carbamothioyl]amino}propanoic    acid;-   (2S)-2-amino-3-{[(3-chloro-4-methyl-5-sulfophenyl)carbamothioyl]amino}propanoic    acid;-   3-[(4S)-4-amino-4-(hydroxycarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonic    acid;-   3-[(4S)-4-amino-4-(hydroxycarbamoyl)butanamide]-5-chloro-4-methylbenzene-1-sulfonic    acid; and-   (2S)-2-amino-3-{[(3-sulfophenyl)carbamoyl]amino}propanoic acid.

Hereinafter, a method for producing compound (I) will be described.

A compound represented by General Formula (I) of the present inventioncan be produced, for example, by the following method.

In the following reaction formula, R¹-R⁶, R^(a), R^(b), R^(c), X, Y, Gand Q are the same groups as those in the definition for theabove-described Formula (I). In addition, in Formula (2), M represents afunctional group that binds to XH or a compound represented by Formula(3) to form X—Y.

For example, when X—Y of Formula (I) represents —CR^(a)R^(b)—CO— groupin Reaction Formula (A), the compound can be produced as follows.

A carboxylic acid derivative (2a) and an amine derivative (3) aredissolved or suspended in an appropriate solvent, and mixed with acondensing agent such as dicyclohexylcarbodiimide (DCC),1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) orN,N-carbonyl diimidazole (CDI) in the presence or absence of a base suchas triethylamine or pyridine, while the reaction system may be cooled,heated or the like as appropriate to produce (I-a). Upon condensation,an adjuvant, a catalyst or the like that adjusts the reaction, such as1-hydroxybenzotriazole (HOBt), 1-hydroxy-7-azabenzotriazole (HOAt) maybe added. The protective group may be removed as appropriate to producean amide derivative represented by General Formula (I-a).

Alternatively, a carboxylic acid halide (2a′) and an amine derivative(3) are mixed in an appropriate solvent and a catalyst such as4-dimethylaminopyridine is used in the presence of a base such astriethylamine or pyridine, while cooling, heating or the like may beperformed as appropriate to produce an amide derivative (I-a). Here, Halin Reaction Formula B2 represents a halogen atom.

In addition, when X—Y of Formula (I) represents —O—CO— group in ReactionFormula A, the compound may be produced as follows. An alcoholderivative (2b) and an amine derivative (3) are dissolved or suspendedin an appropriate solvent, and mixed with a condensing agent such asCDI, phosgene, triphosgene or the like in the presence or absence of abase such as triethylamine or pyridine, while the reaction system may becooled, heated or the like as appropriate to produce a carbamatederivative (I-b).

In addition, a thiocarbonylating agent such as2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiaziphosphetane-2,4-disulfide(Lawesson's reagent) may be reacted with (1-b) in an appropriate solventwith or without heating to produce an O-substituted thiocarbamatederivative (I-b′).

Moreover, in the case where X—Y of Formula (I) in Reaction Formula Arepresents —NR^(c)—CO— group, the compound may be produced, for example,as follows. The amine of an alkylamine derivative represented by (2c) ora salt thereof and the amine derivative of (3) are dissolved orsuspended in an appropriate solvent, and mixed with a condensing agentsuch as CDI, phosgene or triphosgene or a carbonyl source such asdimethyl carbonate in the presence or absence of a base such astriethylamine or pyridine, while the reaction system may be cooled orheated as appropriate to produce an urea derivative (I-c).

Alternatively, as shown in Reaction Formula D2, a method using Lossenrearrangement may be employed as described in Org. Lett., Vol. 11, No.24, 2009, 5622-5625 while using a hydroxamic acid derivative (4) and anamine derivative (3) to produce an urea derivative (I-c′).

Alternatively, the compound may also be produced by a method in which acarbamate derivative (3a) is produced and then substituted with an aminederivative (2c).

Alternatively, in the case where X—Y of Formula (I) represents —S—CO—group in Reaction Formula A, the compound may be produced as follows. Athiol derivative (2d) and an amine derivative of (3) are dissolved orsuspended in an appropriate solvent, and mixed with a condensing agentsuch as CDI, phosgene or triphosgene or a carbonyl source such asdimethyl carbonate in the presence or absence of a base such astriethylamine or pyridine, while the reaction system may be cooled orheated as appropriate to produce a S-substituted thiocarbamatederivative (I-d).

In the case where X—Y of Formula (I) represents N—C(═S)— group inReaction Formula A, the compound may be produced, for example, asfollows. Specifically, an amine derivative represented by (3b) isdissolved or suspended in an appropriate solvent and mixed, for example,with thiophosgene, carbon disulfide or the like in the presence of abase such as sodium carbonate, triethylamine or sodium ethoxide togenerate isothiocyanate (5) as an intermediate. The isothiocyanate (5)may also be a commercially available product. The isothiocyanate, eitherisolated or not isolated, is dissolved in an appropriate solvent, andmixed with amine (2c) of the alkylamine derivative in the presence orabsence of a base such as triethylamine, pyridine or sodium carbonatewhile cooling or heating as appropriate to produce a thiourea derivative(I-e).

The amine derivative (3) may be a commercially available compound or maybe produced as follows.

For example, the following nitro derivative (4) can be used, forexample, with hydrogen gas as a reductant to perform hydrogenationreaction in the presence of a catalyst, or used to perform generalreduction reaction, for example, through reaction with tin chloride toproduce (3b).

In addition, (3c) can be produced by selecting an appropriate reductantupon reduction as described in Journal of the Chemical Society, PerkinTransactions 1: Organic and Bio-Organic Chemistry (1972-1999), 1998, #3,p. 509-520.

Examples of the substituent R⁷ on G include a reaction in which aromatichalide (7) is reacted with a strong base such as n-butyllithium toperform lithium-halogen exchange for reaction with dry ice to introducea carboxyl group, and a reaction in which aromatic halide (7) is reactedwith a transition metal such as palladium for reaction with carbonmonoxide gas. Similarly, a phosphate ester substitute can be produced,for example, by a method described in Synthesis, 1981, #1 p. 56-57 inwhich aromatic halide is reacted with a transition metal catalyst suchas palladium for reaction with phosphite ester, or a sulfonic acidderivative is produced by reacting fuming sulfuric acid with aromaticcompound (8) as shown in Reaction Formula J. Here, in reaction formulaI, Hal, V and Prot represent a halogen atom or a pseudohalogen atom, aprotected nitrogen atom or a group that can change into a nitrogen atomand a protective group or a hydrogen atom, respectively.

Heteroaryls may be synthesized, for example, according to the methoddescribed in Chemical reactivity of aromatic hetero ring compound andring synthesis (Sakamoto et al., Kodansha Scientific). For example, thefollowing heteroaryl can be synthesized by treating a diacylhydrazinederivative with an appropriate dehydrating agent.

A solvent used for the reaction of each of the above-described steps isnot particularly limited as long as it does not interfere with thereaction and dissolves at least a part of the starting material,examples being:

Aliphatic hydrocarbons: hexane, cyclohexane, petroleum ether;

Aromatic hydrocarbons: benzene, toluene, xylene;

Amides: dimethylformamide, N-methyl-2-pyrolidone, dimethylacetamide;

Amines: triethylamine, diisopropylethylamine, pyridine, 2,6-lutidine;

Alcohols: methanol, ethanol, 1-propanol, 2-propanol, 1-butanol,2-butanol;

Ethers: diethylether, dioxane, tetrahydrofuran, dimethoxyethane;

Ketones: acetone, methyl ethyl ketone, methyl isobutyl ketone,cyclohexanone;

Esters: ethyl acetate, isopropyl acetate, butyl acetate;

Acids: formic acid, acetic acid, propionic acid, trifluoroacetic acid,sulfuric acid;

Sulfoxides: dimethylsulfoxide, sulfolane;

Nitriles: acetonitrile, propionitrile;

Halogenated hydrocarbons: dichloromethane, chloroform, carbontetrachloride, dichloroethane, chlorobenzene;

Others: water; and

mixtures thereof.

In each of the above-described steps, it may be preferable to introducea protective group into the functional group of the formulae in advance.As the protective group, for example, functional groups described inProtective Groups in Organic Synthesis, 4th Ed. (WILEY-INTERNATIONAL,WUTS, GREEN) and the like may be used, although the present invention isnot limited thereto. A compound of interest can be obtained byappropriately performing protection and deprotection according to themethod described in the above-mentioned document or the like.

The compound represented by General Formula (I) or a salt thereofproduced as described above can be isolated/purified by knownseparation/purification means such as extraction, condensation, vacuumcondensation, solvent extraction, crystallization, recrystallization,re-extraction, various chromatographies or the like.

The alkylamine derivatives used with the present invention also comprisea form of salt. When the alkylamine derivative of the present inventiontakes a form of salt, the salt should be a pharmaceutically acceptablesalt or an edible salt. For acid groups such as a carboxyl group in theformula, examples of salts include ammonium salt, salts with alkalimetals such as sodium and potassium, salts with alkali earth metals suchas calcium and magnesium, aluminum salt, zinc salt, salts with organicamines such as triethylamine, ethanolamine, morpholine, pyrrolidine,piperidine, piperazine and dicyclohexylamine, and salts with basic aminoacids such as arginine and lysine. For basic groups in the formula, ifany, examples of salts include salts with inorganic acids such ashydrochloric acid, sulfuric acid, phosphoric acid, nitric acid andhydrobromic acid, salts with organic carboxylic acids such as aceticacid, trifluoroacetic acid, citric acid, benzoic acid, maleic acid,fumaric acid, tartaric acid, succinic acid, tannic acid, butyric acid,hibenzic acid, pamoic acid, enanthic acid, decanoic acid, teoclic acid,salicylic acid, lactic acid, oxalic acid, mandelic acid and malic acid,and salts with organic sulfonic acids such as methanesulfonic acid,benzenesulfonic acid and p-toluenesulfonic acid. These salts areproduced by bringing the compound into contact with an acid or a basethat can be used for producing a pharmaceutical product.

According to the present invention, a compound represented by Formula(I) or a salt thereof may be an anhydride or may form a solvate such asa hydrate or an alcohol adduct. The term “solvation” as used hereinrefers to a phenomenon where a solute molecule or ion strongly attractsa solvent molecule adjacent thereto in a solution and form a molecularpopulation. For example, if the solvent is water, it is referred to ashydration. The solvate may be either a hydrate or a non-hydrate. As anon-hydrate, an alcohol (for example, methanol, ethanol, n-propanol),dimethylformamide or the like can be used.

In addition, a compound of the present invention or a salt thereof maybe present in several tautometric forms, for example, enol and imineforms, keto and enamine forms, or a mixture thereof. Tautomers arepresent as a mixture of a tautometric set in a solution. In a solidform, one tautomer is generally dominant over the other. Although onlyone tautomer may be described, the present invention comprises anytautomer of the compound of the present invention.

The present invention comprises all of the stereoisomers (for example,enantiomers, diastereomers (including cis and trans geometric isomers))of the compound represented by Formula (I), racemic forms of theseisomers, and other mixtures. For example, a compound represented byFormula (I) of the present invention may have one or more asymmetriccenters, and the present invention comprises a racemic mixture, adiastereomer mixture and an enantiomer of such a compound.

When a compound according to the present invention is obtained in a formof a free form, it may be converted into a state of a salt formable bythe compound, a hydrate thereof or a solvate thereof according to aroutine method.

On the other hand, when a compound according to the present invention isobtained as a salt, a hydrate or a solvate of the compound, it may beconverted into a free form of the compound according to a routinemethod.

The present invention comprises any isotope of the compound representedby Formula (I). An isotope of a compound of the present invention has atleast one atom substituted with an atom that has the same atomic number(proton number) but has a different mass number (sum of the numbers ofprotons and neutrons). Examples of isotopes included in the compound ofthe present invention include a hydrogen atom, a carbon atom, a nitrogenatom, an oxygen atom, a phosphorus atom, a sulfur atom, a fluorine atomand a chlorine atom, including ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸O, ³¹P,³²P, ³⁵S, ¹⁸F and ³⁶Cl. In particular, unstable radioactive isotopessuch as ³H and ¹⁴C, that release radiation and emit neutron are usefulfor a body tissue distribution test of a pharmaceutical product or acompound. Since a stable isotope does not undergo decay, has almost nochange in the abundance and has no radioactivity, it can be used safely.An isotope of a compound of the present invention may be convertedaccording to a routine method by replacing the reagent used forsynthesis with a reagent containing a corresponding isotope.

A compound of the present invention can be used as a pharmaceuticalagent, particularly as a CaSR agonistic agent, and can be used as aprophylactic or therapeutic agent for a disease that is amelioratedthrough CaSR activation.

CaSR is expressed in various tissues and involved in variousphysiological actions. CaSR senses an increase in the blood calciumlevel in parathyroid, and suppresses secretion of parathyroid hormone(PTH) to correct the blood calcium level. Accordingly, in addition tothe above-mentioned hyperparathyroidism, a compound that activates CaSRis also expected to serve as a therapeutic drug for various diseasessuch as bone diseases and upper and lower digestive disorders (TheJournal of Clinical Investigation, 1997, Vol. 99, p. 2328-2333 and TheAmerican Journal of Physiology-Gastrointestinal and Liver Physiology,2002, Vol. 283, p. G240-G250), diabetes (The Journal of BiologicalChemistry, 1999, Vol. 274, p. 20561-20568 and The Journal of BiologicalChemistry, 2000, Vol. 275, p. 18777-18784), and anterior pituitaryhypofunction/hyperfunction (Molecular Endocrinology, 1996, Vol. 10, p.555-565).

In addition to calcium modulation, CaSR is reported to be expressed inboth mature and undifferentiated adipocytes and involved in differentialinhibition in the adipocytes (Endocrinology. 2005 May; 146(5): 2176-9,Exp Cell Res. 2004 Dec. 10; 301(2):280-92.), expressed in erythroblasts,megakaryocytes and platelets and involved in hematopoiesis regulation inthe myeloid cells (J Bone Miner Res. 1997 December; 12(12):1959-70.),and expressed in gastric parietal cells and involved in gastric acidsecretion (J Clin Endocrinol Metab. 2005 March; 90(3):1489-94).Additionally, CaSR is also reported to be expressed in the followingtissues and involved in the functional regulations thereof: duodenum,jejunum and ileum (Am J Physiol Gastrointest Liver Physiol. 2002 July;283(1): G240-50.), large intestine (Am J Physiol Gastrointest LiverPhysiol. 2002 July; 283(1): G240-50.), epidermal keratinocytes (CellCalcium. 2004 March; 35(3): 265-73.), hepatocytes (J Biol. Chem. 2001Feb. 9; 276(6): 4070-9.), epithelium lentis (Biochem Biophys Res Commun.1997 Apr. 28; 233(3): 801-5.), pancreatic Langerhans' islet β cells(Endocrine. 1999 December; 11(3): 293-300.), lung (J Clin EndocrinolMetab. 1998 February; 83(2): 703-7.), monocytic cells (J Clin Invest.2000 May; 105(9): 1299-305.), osteoblasts (Endocrinology. 2004 July;145(7): 3451-62, Am J Physiol Endocrinol Metab. 2005 March; 288(3):E608-16. Epub 2004 Nov. 16.) and the like.

Moreover, since glutathione, which is known as an agent for impartingkokumi, has been confirmed to show a calcium receptor activating effect,and that a peptide derivative having a CaSR agonistic activity presentskokumi (WO2007/055393), a compound having a CaSR agonistic activity issuggested to be useful as an agent for imparting kokumi.

In particular, calcium receptors are expressed in the G cells and theparietal cells of the stomach, and are found to have an effect ofstimulating gastrin and gastric acid secretion (Journal of ClinicalInvestigation (1997), 99: 2328-2333, Gastroenterology 1999; 116:118-126). In addition, calcium receptors are expressed in the largeintestine and regulates secretion of water (The American Journal ofPhysiology-Gastroinstinal and Liver Physiology (2002), 283: G240-G250).Since calcium receptor agonists such as cinacalcet and gamma-glutamylpeptide derivatives have been shown to have an effect of suppressingdiarrhea in animal models (WO2008/139947), an effect of stimulatingbicarbonic acid or somatostatin secretion, and an effect of reducing aninjury area in non-steroidal anti-inflammatory drug (NSAID)-inducedsmall intestine inflammation animal models (WO2009/119554), a compoundhaving a CaSR agonistic effect is found beneficial as a prophylactic ortherapeutic agent for diarrhea or acid secretion-associated diseasessuch as, gastric ulcer, duodenal ulcer and reflux esophagitis as well asan appetite-modulating agent.

Furthermore, since a peptide or a low-molecular compound with calciumreceptor activation has been shown to stimulate secretion of GLP-1 andCCK in intestinal tract-derived STC-1 and GLUTag cells (WO2009/11221), acompound having a CaSR agonistic effect is found beneficial as aprophylactic or therapeutic agent for diabetes and obesity.

Moreover, since cinacalcet and gamma-glutamylvaline have been confirmedto have an IgA production-promoting ability through LPS stimulation andan IgG production-promoting effect through ConA stimulation(WO2009/128523), a compound having a CaSR agonistic effect is foundbeneficial as an immunostimulator or as a therapeutic or prophylacticagent for a disease that is effectively prevented or treated byimmunostimulation, for example, various infectious diseases, diarrhea,polyp, tumor, enteritis or allergy.

Accordingly, a compound of the present invention can be used as anactive ingredient of a pharmaceutical composition for preventing ortreating a disease that is ameliorated through CaSR activation.

Here, “a disease that is ameliorated through CaSR activation” is anillness or deficiency characterized by abnormal calcium homeostasis, oran illness or a condition that is induced by reduction in CaSR function,specific examples being diarrhea, diseases associated with secretion ofdigestive tract acid, eating disorders such as excessive appetite,hyperparathyroidisms (primary and secondary parathyroid hyperfunctions,and secondary hyperparathyroidism under maintenance dialysis), diabetes,obesity, compromised immune function, Paget's disease, malignanthypercalcemia, osteoporosis and hypertension.

The diseases associated with secretion of digestive tract acid includeulcer and inflammatory diseases in digestive tracts such as the stomachor the small intestine (duodenum, jejunum, ileum), which include thoseinduced by endogenous causes such as stress or the like, and thoseinduced by exogenous causes such as drugs (non-steroidalanti-inflammatory drugs, alcohol or the like).

Examples of “peptic ulcer” include gastric ulcer, duodenal ulcer,gastritis, NSAID-induced small intestine inflammation, refluxesophagitis, non-erosive gastroesophageal reflux disease andnon-steroidal anti-inflammatory drug-induced ulcer.

The compound of the present invention is administered directly or as apharmaceutical composition that contains the compound of the presentinvention as an active ingredient. A method for applying such apharmaceutical composition is not particularly limited, and oraladministration, invasive administration using injection or the like,suppository administration or transdermal administration may beemployed. The active ingredient can be mixed with a non-toxic solid orliquid carrier for a pharmaceutical agent appropriate for the givenmethod such as oral administration, injection or the like, andadministered in a form of a common pharmaceutical formulation. Examplesof such formulations include formulations in a solid form such as atablet, granules, a pill, powder, a capsule, a suppository, asugarcoated tablet or a depot formulation, liquid formulation such as asolution formulation, suspension and emulsion, and a lyophilizedformulation. These formulations can be prepared by pharmaceuticallycommon means.

Examples of the above-described non-toxic carrier for a pharmaceuticalagent include glucose, lactose, sucrose, starch, calcium carbonate,calcium phosphate, mannitol, dextrin, fatty acid glyceride, polyethyleneglycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitanfatty acid ester, gelatin, albumin, amino acid, water and physiologicalsaline. If necessary, a common additive such as a stabilizer, amoisturizer, an emulsifier, a binder or a tonicity agent mayappropriately be added.

A compound of the present invention can also be used as eatables thathave an effect of treating or preventing a CaSR-involved disease. Forexample, it may be made into eatables with a container or a packagethereof indicating that it has an effect of treating or preventing theabove-mentioned CaSR-involved disease.

A dosage form and an administration form of a compound of the presentinvention are not particularly limited, and it may be given by oraladministration or by parenteral administration (intake) such asadministration by intravenous drip, or administration by injection(transvenous administration). For the sake of easier administration,oral administration is favorable but the administration is not limitedthereto.

For an orally administered agent, granules, fine granules, a powderedagent, a coated tablet, a tablet, a suppository, powder, a (micro)capsule, a chewable agent, syrup, juice, a liquid agent, suspension andemulsion can be employed. For an injectable agent, those for directintravenous infusion, those for administration by intravenous drip, andformulation that prolongs the release of the active substance can beemployed. Thus, a dosage form of a general pharmaceutical formulationscan be employed.

In the case of oral administration, the dosage differs depending on thecondition and age of the patient as well as the given method, but itshould be an amount effective for treatment or prevention, which mayappropriately be adjusted according to the age, sex, weight, conditionand the like of the patient. For example, in the case of an oraladministration, an amount of a CaSR agonist per day, in general, ispreferably 0.001 mg-10 mg and more preferably 0.1 mg-1 mg per kilogramweight of an adult.

Moreover, a dosage, in the case of parenteral administration such asadministration by intravenous drip or administration by injection(transvenous administration), is preferably about one-tenth toone-twentieth of the above-described range of preferable dosage (amountof intake) for oral administration.

The above-described dosage for oral administration may similarly beapplied to the later-described food, which does not preclude that theCaSR agonist is contained in the food such that the amount of intake islower than that for administration.

A compound of the present invention can be formulated according to aroutine method. According to the requirement for the formulation,various pharmacologically acceptable formulation substances (such asadjuvant) can be blended. A formulation substance can appropriately beselected according to the dosage form of the formulation, examples beingan excipient, a diluent, an additive, a disintegrant, a binder, acoating agent, a lubricant, a sliding agent, a lubricator, a flavoringagent, a sweetening agent and a solubilizer. Furthermore, specificexamples of the formulation substance include magnesium carbonate,titanium dioxide, lactose, mannitol and other sugars, talc, milkprotein, gelatin, starch, cellulose and derivatives thereof, animal andplant oils, polyethylene glycol, solvents such as sterile water andmono- or polyalcohol such as glycerol.

Other than a routine method, a compound of the present invention canalso be formulated according to various pharmaceutical formulation formsthat will be developed in the future. For such formulations, methodsdeveloped in the future can appropriately be employed.

A package containing a compound of the present invention may includedirections providing explanations about use thereof. The directions maybe, for example, a so-called package insert that provides explanatorymatter related to use, efficacy, an administration method and the like.

A compound of the present invention may be contained in food. The formof food is not particularly limited, and can be produced by the sameproduction method and with the same materials as general food exceptthat a CaSR agonist is blended therein. Examples of food includeseasonings; beverage such as juice and milk; sweets; jelly; health food;processed agricultural products; processed seafood products; processedlivestock products such as milk; and supplementary food. In addition,such food can be provided as food with health claims, including foodlabeled with a claim that it is used for preventing, treating orameliorating acid secretion-associated diseases, in particular, food forspecified health use.

When a compound of the present invention is used as supplementary food,it may be prepared into a form such as a tablet, a capsule, powder,granule, suspension, a chewable agent or syrup. Other than those takenin as food, supplementary food according to the present invention alsorefers to those taken in for the purpose of supplying nutrition,including nutritious supplements, supplements and the like. In addition,the supplementary food of the present invention also includes some ofthe food with health claims.

A method for using an agent for imparting kokumi that contains one ormore types of compounds selected from the compounds of the presentinvention as active ingredients is not particularly limited, and it maybe used by adding to eatables such as seasonings, food, beverage or thelike.

A substance for imparting kokumi of the present invention may be usedalone or in combination with other various additives or the like to beused by being added to eatables such as seasonings, food and beverage.

Moreover, an agent for imparting kokumi of the present invention mayconsist, for example, only of one or more types of compounds selectedfrom the above-described compounds of the present invention, or it mayfurther be added with an existing compound having kokumi impartingactivity (glutathione, alliin, etc.), various additives and the like. Inthis respect, one or more types of existing compounds with CaSRstimulating activity may be added, and the present invention alsocomprises such compounds.

Herein, “imparting kokumi” refers to enhancement of any one of the fivebasic tastes, i.e., sweetness, saltiness, sourness, bitterness and umami(savory taste), and impartment of tastes associated with the basictastes such as richness, thickness, growth (or mouthfulness), continuityand harmony that come along with the basic tastes. Moreover, an agentfor imparting kokumi can also be expressed as a flavor enhancer. Hence,an agent for imparting kokumi of the present invention can also be usedas a sweetness enhancer, a saltiness enhancer, a sourness enhancer, abitterness enhancer or a umami enhancer.

Examples of existing compounds with CaSR activity include cations suchas calcium and gadolinium, basic peptides such as polyarginine andpolylysine, polyamines such as putrescine, spermine and spermidine,proteins such as protamine, peptides such as phenylalanine andglutathione, and cinacalcet. These compounds may also take a form ofacceptable salt.

The above-mentioned additives can be used without particular limitationas long as they are known to be able to be added to and blended intoeatables such as seasonings, food and beverage. Examples of suchadditives include flavors, sugars, sweeteners, food fiber, vitamins,amino acids such as monosodium glutamate (MSG), nucleic acids such asinosine monophosphate (IMP), inorganic salts such as sodium chloride,and water.

An amount of a substance for imparting kokumi of the present inventionor an agent for imparting kokumi of the present invention used foreatables should be an effective amount for imparting kokumi, and canappropriately be adjusted according to use. For example, in the case ofseasonings, food or beverage, a total amount of an agent for impartingkokumi or a substance for imparting kokumi of the present invention inthe seasonings, food or beverage is 1 wt·ppb-99.9 wt %, preferably 10wt·ppb-99.9 wt %, and more preferably about 10 wt·ppm-10 wt %.

Thus, one or more types of the substances for imparting kokumi of thecompound of the present invention or the agents for imparting kokumi ofthe present invention can be added to eatables to give a content of 1wt·ppb-99.9 wt %, preferably 10 wt·ppb-99.9 wt %, and more preferablyabout 10 wt·ppm-10 wt % so as to produce eatables with an impartedkokumi.

Furthermore, the above-described seasonings that has been imparted withkokumi by containing 1 wt·ppb-99.9 wt % of one or more types of thesubstances for imparting kokumi of the present invention or the agentsfor imparting kokumi of the present invention can be added to eatablesto give a content of 0.01-10 wt % and preferably 0.1-10 wt % so as toproduce eatables with an imparted kokumi.

A form of the substance for imparting kokumi of a compound of thepresent invention or the agent for imparting kokumi of the presentinvention, which is added to eatables is not limited in terms of itsphysicality, i.e., whether dry powder, paste, solution or the like.

EXAMPLES

The present invention will be described in detail by means of examplesbelow. They are preferable embodiments of the present invention and thepresent invention should not be limited to these examples. Thestructures and MS value or NMR measurements of the compounds synthesizedaccording to the following methods are shown in Tables 1-15.

In these examples, purification step A refers to a step of lyophilizinga fraction of interest by performing elution with a mixed solution ofwater and acetonitrile containing 0.1% trifluoroacetic acid (v/v) upon areversed-phase high-performance liquid chromatography that uses a silicagel chemically bounded with an octadodecyl group as a filler. A routinemethod generally refers to a synthetic chemical method, for example,solvent extraction, back extraction, washing, neutralization and drying.

Example 1 Synthesis of (2S)-2-amino-4-[(pyridine-2-yl)carbamoyl]butanoicacid trifluoroacetic acid salt

70 mg (0.188 mmol, 1 equivalent) of Cbz-Glu-OBzl and 85 mg (0.226 mmol)of O-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU) were dissolved in 1 ml of DMF, added with 39μl (0.282 mmol) of triethylamine and stirred for 5 minutes. 18 mg of2-amino-pyridine was added, and stirred at room temperature overnight.Aftertreatment was performed according to a routine method and theresulting crude product was dissolved in 3 ml of acetic acid, to which acatalyst amount of palladium carbon (Pd/C) was added and stirred in ahydrogen atmosphere overnight. After filtrating the catalyst, thesolvent was distilled away, and the resulting residue was subjected topurification step A to obtain the title compound.

Yield: 23.1 mg

Example 2 Synthesis of(2S)-2-amino-4-[(5-bromo-6-methylpyridine-2-yl)carbamoyl]butanoic acidtrifluoroacetic acid salt

100 mg (0.33 mmol) of Boc-Glu-OtBu, 53.8 mg (0.40 mmol) of1-hydroxy-7-azabenzotriazole and 150.4 mg (0.40 mmol) ofO-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate were dissolved in 1 ml of DMF, added with 68.5 μl(0.49 mmol) of triethylamine and stirred at room temperature for 5minutes. 61.7 mg (0.33 mmol) of 6-amino-3-bromo-2-methylpyridine wasadded and stirred at room temperature overnight. After diluting thereaction solution with water and acetonitrile, the purification step Awas used to obtain 109.3 mg of the crude purified substance of the titlecompound in protected form. To the obtained crude purified substance, 2ml of trifluoroacetic acid was added and the resultant was stirred atroom temperature overnight. After distilling the solvent away, theresultant was purified using purification step A to obtain the titlecompound.

Yield: 43.36 mg

Example 3 Synthesis of (2S)-2-amino-4-[(3-sulfophenyl)sulfamoyl]butanoicacid

According to the method described in J. Med. Chem. 1999, 42, 5197-5211,0.24 g (0.425 mmol) of2-{[(benzyloxy)carbonyl]amino}-4-[(3-{[(benzyloxy)carbonyl]amino}-4-methoxy-4-oxobutyl)disulfanyl]methylbutanoate was obtained as an intermediate. To 0.24 g of the resultingintermediate, 2 ml of acetic acid and 0.5 ml of water were added and 87μl (1.7 mmol) of bromine was added while cooling with ice. Afteragitation for 20 minutes, the solvent was distilled away, andaftertreatment was performed with ethyl acetate and 1M hydrochloric acidaccording to a routine method to distill away the solvent. To theresulting residue, 191 mg (1.1 mmol) of 3-aminobenzenesulfonic acid wasadded and the resultant was suspended in 5 ml of methylene chloride. Tothis, 0.52 ml (3 mmol) of N,N-diisopropylethylamine was added andstirred overnight. After distilling the solvent away, purification stepA was used to obtain 33.2 mg of the crude product.

The resulting crude product was dissolved in 1 ml of tetrahydrofuran,0.5 ml of methanol and 0.5 ml of water, to which 9 mg of lithiumhydroxide was added. Following an hour of agitation, the solvent wasdistilled away. To the resulting residue, 3 ml of 48% hydrogenbromide-acetic acid solution was added and stirred for an hour. Afterdistilling the solvent away, purification step A was used to obtain thetitle compound.

Yield: 9.2 mg

Example 4 Synthesis of(2R)-2-amino-3-{[(3-sulfophenyl)carbamoyl]sulfanyl}propanoic acid

To 171 mg (1 mmol) of L-cysteine methyl ester hydrochloride, 2 ml oftetrahydrofuran and 0.3 ml of triethylamine were added, 218 mg (1 mmol)of di-tert-butyl dicarbonate dissolved in 2 ml of tetrahydrofuran wasadded and the resultant was stirred at room temperature for 2 hours.Ethyl acetate and 10% aqueous citric acid solution were used foraftertreatment according to a routine method to obtain a crude productof Boc-Cys-OMe. To the obtained crude product, 191 mg of 3-aminobenzenesulfonic acid and 100 mg (0.33 mmol) of triphosgene were added and theresultant was suspended in 3 ml of methylene chloride. To this, 3 mmolof N,N-diisopropylethylamine was added and stirred for 2 hours. Afterdistilling the solvent away, the resultant was diluted with water andacetonitrile, and purification step A was employed to obtain 72 mg of acrude purified substance of the title compound in protected form. 72 mgof the resulting crude purified substance was dissolved in 1 ml oftetrahydrofuran, 0.5 ml of methanol and 0.5 ml of water, to which 17 mgof lithium hydroxide was added and the resultant was stirred at roomtemperature for 2 hours. After distilling the solvent away, 2 ml oftrifluoroacetic acid was added and stirred for 2 hours. The resultantresidue was purified by purification step A to obtain the titlecompound.

Yield: 44.0 mg

Example 5 Synthesis of(2R)-2-amino-3-{[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]sulfanyl}propanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 4 was replaced with3-amino-5-chloro-2-hydroxybenzenesulfonic acid.

Yield: 15.0 mg

Example 6 Synthesis of(2S)-2-amino-3-{[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]amino}propanoicacid

297 mg (1 mmol) of 3-amino-N-(tert-butoxycarbonyl)-L-alaninetert-butylester hydrochloride, 223 mg (1 mmol) of3-amino-5-chloro-2-hydroxybenzenesulfonic acid and 100 mg (0.33 mmol) oftriphosgene were suspended in methylene chloride (3 ml), added with 0.8ml of pyridine, and stirred at room temperature overnight. The solventwas distilled away, and the resultant residue was purified by employingpurification step A to obtain the title compound in protected form. Tothe resulting protected form, 3 ml of trifluoroacetic acid was added,and the resultant was stirred for 5 hours. Subsequently, the solvent wasdistilled away and the obtained residue was purified using purificationstep A to obtain the title compound.

Yield: 20.1 mg

Example 7 Synthesis of3-({[(2S)-2-amino-propoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

75 mg (1 mmol) of (S)-2-(tert-butoxycarbonylamino)-1-propanol(Boc-Ala-ol), 223 mg (1 mmol) of3-amino-5-chloro-2-hydroxybenzenesulfonic acid and 100 mg (0.33 mmol) oftriphosgene were suspended in 3 ml of methylene chloride, and added with0.7 ml of pyridine. Following agitation at room temperature overnight,the solvent was distilled away, purified using purification step A toobtain a crude purified substance of the title compound in protectedform. To the resultant crude purified substance, 2 ml of trifluoroaceticacid was added, and the resultant was stirred at room temperature for 2hours. Subsequently, the solvent was distilled away, and the resultingresidue was added with water and acetonitrile for deposition and thedeposited solid was filtrated to obtain the title compound.

Yield: 140.1 mg

Example 83-({[(2S)-2-amino-3-methylbutoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatBoc-Ala-ol used in Example 7 was replaced with(S)-(−)-2-(butoxycarbonylamino-3-methyl 1-butanol (Boc-Val-ol).

Yield: 108.5 mg

Example 9 Synthesis of3-({[(2S)-2-amino-3-phenylpropoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

285 mg (1 mmol) of (S)-2-(benzyloxycarbonylamino)-3-phenyl-1-propanol,223 mg (1 mmol) of 3-amino-5-chloro-2-hydroxybenzenesulfonic acid and100 mg (0.33 mmol) of triphosgene were suspended in 3 ml of methylenechloride, to which 0.7 ml of pyridine was dropwisely added. Followingagitation at room temperature overnight, the solvent was distilled away,and the resultant was purified using purification step A to obtain acrude purified substance of the title compound in protected form. To theobtained crude purified substance, 2 ml of 48% hydrogen bromide-aceticacid solution was added, and the resultant was stirred at roomtemperature for 2 hours. The solvent was distilled away and water andacetonitrile were added to the obtained residue for deposition. Thedeposited solid substance was filtrated to obtain the title compound.

Yield: 151.6 mg

Example 10 Synthesis of3-({[(2S)-2-amino-2-phenylethoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatBoc-Ala-ol used in Example 7 was replaced with(S)—N-(tert-butoxycarbonyl)-2-phenylglycinol.

Yield: 55.2 mg

Example 11 Synthesis of3-({[(2S)-2-amino-4-carbamoylbutoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatBoc-Ala-ol used in Example 7 was replaced with (S)-tert-butyl5-amino-1-hydroxy-5-oxopentane-2-ylcarbamate (Boc-Gln-ol).

Yield: 25.2 mg

Example 12 Synthesis of3-({[(2S)-2-amino-4-cyanobutoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

The compound was obtained as a by-product during synthesis in Example11.

Yield: 6.2 mg

Example 13 Synthesis of(2S)-2-amino-3-{[(3-chloro-4-methyl-5-sulfophenyl)carbamoyl]amino}propanoicacid

100 mg (0.336 mmol) of 3-amino-N-(tert-butoxycarbonyl)-L-alaninetert-butylester hydrochloride, 74 mg (0.336 mmol) of5-amino-3-chloro-2-methylbenzenesulfonic acid and 55 mg (0.336 mmol) ofN,N-carbonyl diimidazole were added with 1 ml of methylene chloride and0.25 ml of pyridine, and the resultant was stirred at room temperatureovernight. The solvent was distilled away, and the resulting residue waspurified using purification step A to obtain the title compound inprotected form. The obtained protected form was added with 3 ml oftrifluoroacetic acid, stirred for 2 hours, and then the solvent wasdistilled away. The resulting residue was purified using purificationstep A to obtain the title compound.

Yield: 43.98 mg

Example 14 Synthesis of(2S)-2-amino-3-{[(3-chloro-2-methyl-5-sulfophenyl)carbamoyl]amino}propanoicacid

The title compound was obtained through a similar operation except that3-amino-5-chloro-4-methylbenzenesulfonic acid was used instead of5-amino-3-chloro-2-methylbenzenesulfonic acid in Example 13.

Yield: 4.0 mg

Example 15 Synthesis of3-[(4S)-4-amino-4-(5-methyl-1,3,4-oxadiazol-2-yl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid Step 1 Synthesis of(4S)-4-{[(tert-butoxy)carbonyl]amino}-4-(5-methyl-1,3,4-oxadiazole-2-yl)butanoicacid benzyl ester

674 mg (2 mmol) of(S)-5-(benzyloxy)-2-(tert-butoxycarbonylamino)-5-oxopentanoic acid(Boc-Glu (OBzl)-OH) and 148 mg (2 mmol) of acetohydrazide were addedwith 5 ml of tetrahydrofuran, added with 356 mg of N,N-carbonyldiimidazole and stirred at room temperature overnight. The solvent wasdistilled away, and the resultant was purified using purifiedpurification step A to obtain a crude purified substance of(4S)-4-(N′-acetylhydrazinecarbonyl)-4-{[(tert-butoxy)carbonyl]amino}butanoicacid benzyl ester. The obtained crude purified substance was dissolvedin 2 ml of methylene chloride, which was added with 72 mg of Burgessreagent and stirred at room temperature overnight. Another 72 mg ofBurgess reagent was added and stirred for two days. After distilling thesolvent away, purification was carried out using purification step A toobtain the title compound.

Step 2

The compound obtained in Step 1 was dissolved in ethyl acetate, addedwith a catalyst amount of Pd/C, and stirred in a hydrogen atmosphereovernight. The catalyst was filtrated and the solvent was distilledaway, thereby obtaining a crude product. To the resulting crude product,6 mg of 3-amino-5-chloro-2-hydroxybenzenesulfonic acid, 4.3 mg of1-hydroxy-7-azabenzotriazole, 12 mg ofO-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate, 0.5 ml of DMF, and 11 μl of triethylamine wereadded and the resultant was stirred overnight. Following dilution withwater and acetonitrile, a crude purified substance that had beenpurified by purification step A was obtained. The obtained crudepurified substance was added with 1 ml of trifluoroacetic acid andstirred for an hour. Thereafter, the solvent was distilled away, and theobtained residue was purified by purification step A to obtain the titlecompound.

Yield: 5.2 mg

Example 16 Synthesis of3-[(4S)-4-amino-4-(5-benzyl-1,3,4-oxadiazol-2-yl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid Step 1 Synthesis of(4S)-4-(5-benzyl-1,3,4-oxadiazole-2-yl)-4-[(tert-butoxy)carbonyl]amino/butanoicacid benzyl ester

337 mg (1 mmol) of Boc-Glu (OBzl)-OH and 150 mg (1 mmol) of2-phenylacetohydrazide were dissolved in methylene chloride, to which235 mg of 1-(3-dimethylamino-propyl)-3-ethylcarbodiimide hydrochloride,183 mg of 1-hydroxybenzotriazole monohydrate and 278 μl of triethylaminewere sequentially added. Following agitation at room temperature for 3hours, ethyl acetate, 1M aqueous sodium hydroxide solution, 1Mhydrochloric acid and saturated saline were used for aftertreatment, andthen the solvent was distilled away to obtain 0.392 g of a crudepurified substance. The obtained crude purified substance was dissolvedin 5 ml of methylene chloride, added with 260 mg of Burgess reagent andstirred for three days. Ethyl acetate, 1M aqueous sodium hydroxidesolution, 1M hydrochloric acid and saturated saline were used foraftertreatment, and then the solvent was distilled away and theresultant was purified by silica gel column chromatography to obtain thetitle compound.

Yield: 96 mg

Step 2

A similar operation to Step 2 in Example 15 was performed on thecompound obtained in Step 1 to obtain the title compound.

Yield: 27.6 mg

Example 17 Synthesis of3-[(4S)-4-amino-5-oxo-5-(2-phenylacetohydrazide)pentanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The compound was obtained as a by-product during the operation inExample 16.

Yield: 44.4 mg

Example 18 Synthesis of3-[(4S)-4-amino-4-(5-benzyl-1,3,4-oxadiazole-2-yl)butanamide]benzene-1-sulfonicacid

In Example 16, the title compound was obtained by performing a similaroperation except that 3-amino-5-chloro-2-hydroxybenzenesulfonic acidused in Step 2 was replaced with 3-aminobenzenesulfonic acid.

Yield: 15.6 mg

Example 19 Synthesis of3-[(4S)-4-amino-4-[5-(4-bromophenyl)-1,3,4-oxadiazole-2-yl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that2-phenylacetohydrazide used in Step 1 in Example 16 was replaced with4-bromobenzhydrazide.

Yield: 48.9 mg

Example 20 Synthesis of3-[(4S)-4-amino-4-[5-(4-bromophenyl)-1,3,4-oxadiazole-2-yl]butanamide]benzene-1-sulfonicacid

The title compound was obtained through a similar operation except that2-phenylacetohydrazide used in Step 1 in Example 16 was replaced with4-bromobenzhydrazide, and 3-amino-5-chloro-2-hydroxybenzenesulfonic acidused in Step 2 was replaced with 3-aminobenzenesulfonic acid.

Yield: 73.2 mg

Example 21 Synthesis of3-[(4S)-4-amino-4-(5-phenyl-1,3,4-oxadiazol-2-yl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained by performing Steps 1 and 2 except thatbenzhydrazide was used instead of 2-phenylacetohydrazide used in Step 1in Example 16.

Yield: 25.6 mg

Example 22 Synthesis of3-[(4S)-4-amino-4-(benzylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

2.0 mmol of Boc-Glu (OBzl)-OH, 2.2 mmol of1-(3-dimethylamino-propyl)-3-ethylcarbodiimide hydrochloride and 2.2mmol of 1-hydroxybenzotriazole monohydrate were dissolved in methylenechloride, to which 2.5 mmol of triethylamine was added. To this, 2.0mmol of benzylamine was added and stirred overnight. Ethyl acetate, 1Maqueous sodium hydroxide solution, 1M hydrochloric acid and saturatedsaline were used for aftertreatment, and then the solvent was distilledaway. The resultant residue was dissolved in 5 ml of THF, 2.5 ml ofmethanol and 2.5 ml of water, added with 90 mg of lithium hydroxide andstirred at room temperature for two hours. Ethyl acetate, 1Mhydrochloric acid and saturated saline were used for aftertreatment, andthen the solvent was distilled away to obtain 440 mg of residue. Theobtained residue was added with 261 mg of 1-hydroxy-7-azabenzotriazole,730 mg of O-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate and 330 mg of3-amino-5-chloro-2-hydroxybenzenesulfonic acid, and the resultant wasdissolved in 5 ml of DMF. To this, 412 μl of triethylamine was added andstirred at room temperature overnight. After distilling the solventaway, the resultant was purified by employing purification step A toobtain a crude product of the title compound in protected form. Theobtained crude product was dissolved in 5 ml of methylene chloride and 3ml of trifluoroacetic acid and stirred for 2 hours. After distilling thesolvent away, the resultant was purified by employing purification stepA to obtain the title compound.

Yield: 76.3 mg

Example 23 Synthesis of3-[(4S)-4-amino-4-[(2-phenylethyl)carbamoyl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

2.0 mmol of Boc-Glu (OMe)-OH, 2.2 mmol of1-(3-dimethylamino-propyl)-3-ethylcarbodiimide hydrochloride and 2.2mmol of 1-hydroxybenzotriazole monohydrate were dissolved in methylenechloride, to which 2.5 mmol of triethylamine was added. To this, 2.0mmol of 2-phenylethylamine was added and stirred overnight. Ethylacetate, 1M aqueous sodium hydroxide solution, 1M hydrochloric acid andsaturated saline were used for aftertreatment, and then the solvent wasdistilled away. The obtained residue was dissolved in 5 ml of THF, 2.5ml of methanol and 2.5 ml of water, to which 90 mg of lithium hydroxidewas added and stirred at room temperature for 2 hours. Ethyl acetate, 1Mhydrochloric acid and saturated saline were used for aftertreatment, andthen the solvent was distilled away to obtain 440 mg of residue. Theobtained residue was added with 261 mg of 1-hydroxy-7-azabenzotriazole,730 mg of O-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate and 330 mg of3-amino-5-chloro-2-hydroxybenzenesulfonic acid, which was dissolved in 5ml of DMF. To this, 412 μl of triethylamine was added and stirred atroom temperature overnight. After distilling the solvent away, theresultant was purified by employing purification step A to obtain acrude product of the title compound in protected form. The obtainedcrude product was dissolved in 5 ml of methylene chloride and 3 ml oftrifluoroacetic acid, and stirred for 2 hours. After distilling thesolvent away, water and acetonitrile were added for deposition and thedeposited solid substance was filtrated to obtain the title compound.

Yield: 151.9 mg

Example 24 Synthesis of3-[(4S)-4-amino-4-[(3-phenylpropyl)carbamoyl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that2-phenylethylamine in Example 23 was replaced with 3-phenylpropylamine.

Yield: 126.7 mg

Example 25 Synthesis of3-[(4S)-4-amino-4-[(5-phenylbutyl)carbamoyl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that2-phenylethylamine in Example 23 was replaced with 4-phenylbutylamine.

Yield: 135.9 mg

Example 26 Synthesis of3-[(4S)-4-amino-4-[5-[4-(dimethylamino)phenyl]-1,3,4-oxadiazole-2-yl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained by carrying out Steps 1 and 2 exceptthat 4-(dimethylamino)benzhydrazide was used instead of2-phenylacetohydrazide used in Step 1 in Example 16.

Yield: 48.6 mg

Example 27 Synthesis of3-[(4S)-4-amino-4-[5-(thiophene-2-yl)-1,3,4-oxadiazolo-2-yl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained by carrying out Steps 1 and 2 exceptthat 2-thiophenecarboxylic acid hydrazide was used instead of2-phenylacetohydrazide used in Step 1 in Example 16.

Yield: 40.1 mg

Example 28 Synthesis of(2S)-2-amino-3-{[(2-hydroxy-3-sulfophenyl)carbamoyl]amino}propanoic acid

The compound obtained in Example 6 was dissolved in water and methanol,to which a catalyst amount of Pd/C was added, and the resultant wasstirred for three days in a hydrogen atmosphere. After filtrating thecatalyst, the solvent was distilled away, and the resultant was purifiedusing purification step A to obtain the title compound.

Yield: 1.2 mg

Example 29 Synthesis of3-{[(2-aminoethyl)carbamoyl]amino}-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thattert-butyl N-(2-aminoethyl)carbamate was used instead of3-amino-N-(tert-butoxycarbonyl)-L-alanine tert-butylester hydrochlorideused in Example 6.

Yield: 1.0 mg

Example 30 Synthesis of3-[(4S)-4-amino-4-{[2-(1H-indole-3-yl)ethyl]carbamoyl}butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thattryptamine was used instead of 2-phenylethylamine used in Example 23.

Yield: 61.9 mg

Example 31 Synthesis of3-[(4S)-4-amino-4-{[2-(3-methoxyphenyl)ethyl]carbamoyl}butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-methoxyphenethylamine was used instead of 2-phenylethylamine used inExample 23.

Yield: 80.8 mg

Example 32 Synthesis of3-[(4S)-4-amino-4-{[2-(3-chlorophenyl)ethyl]carbamoyl}butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-chlorophenethylamine was used instead of 2-phenylethylamine used inExample 23.

Yield: 122 mg

Example 33 Synthesis of3-[(4S)-4-amino-4-{[3-(1H-imidazole-1-yl)propyl]carbamoyl}butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that1-(3-aminopropyl)imidazole was used instead of 2-phenylethylamine usedin Example 23.

Yield: 20 mg

Example 34 Synthesis of3-[(4S)-4-amino-4-(methanesulfonylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

337 mg (1 mmol) of Boc-Glu (OBzl)-OH, 95 mg (1 mmol) ofmethanesulfonamide, 250 mg (1.2 mmol) of dicyclohexylcarbodiimide and acatalyst amount of 4-(dimethylamino)pyridine were added and dissolved in5 ml of methylene chloride. After agitation at room temperatureovernight, the insoluble matter was filtrated and the solution wasdistilled away. Thereafter, ethyl acetate, 1M hydrochloric acid andsaturated saline were used for aftertreatment. The resulting residue wasdissolved in ethyl acetate, to which a catalyst amount of Pd/C wasadded, and the resultant was stirred in a hydrogen atmosphere overnight.After the catalyst was filtrated, the solvent was distilled away toobtain a residue. To the obtained residue, 220 mg of3-amino-5-chloro-2-hydroxybenzenesulfonic acid, then 150 mg of1-hydroxy-7-azabenzotriazole and 400 mg ofO-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate were added, and 1 ml of DMF were added. Followingaddition of 200 μl of triethylamine, agitation took place overnight.After confirming the reaction progress, the reaction solution wasdiluted with water and acetonitrile and purified by purification step A.The resulting crude purified substance was dissolved in 2 ml ofmethylene chloride and 2 ml of trifluoroacetic acid. After two hours ofagitation, the solvent was distilled away and the resultant was purifiedby purification step A to obtain the title compound.

Yield: 15.3 mg

Example 35 Synthesis of3-[(4S)-4-amino-4-(methanesulfonylcarbamoyl)butanamide]benzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid was used instead of3-amino-5-chloro-2-hydroxybenzenesulfonic acid used in Example 34.

Yield: 7.6 mg

Example 36 Synthesis of5-[(4S)-4-amino-4-(methanesulfonylcarbamoyl)butanamide]-3-chloro-2-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that5-amino-3-chloro-2-methylbenzenesulfonic acid was used instead of3-amino-5-chloro-2-hydroxybenzenesulfonic acid used in Example 34.

Yield: 108.4 mg

Example 37 Synthesis of3-[(4S)-4-amino-4-(methanesulfonylcarbamoyl)butanamide]-5-chloro-4-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-amino-5-chloro-4-methylbenzenesulfonic acid was used instead of3-amino-5-chloro-2-hydroxybenzenesulfonic acid used in Example 34.

Yield: 86.8 mg

Example 38 Synthesis of3-[(4S)-4-amino-4-(hydroxycarbamoyl)butanamide]benzene-1-sulfonic acid

337 mg (1 mmol) of Boc-Glu-OBzl, 173 mg (1 mmol) of3-aminobenzenesulfonic acid, and 420 mg (1.1 mmol) ofO-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate were dissolved in 2 ml of DMF, added with 200 μl oftriethylamine and stirred at room temperature overnight. Followingdilution with water and acetonitrile, purification step A was used toobtain a crude purified substance. 50 mg of the obtained crude purifiedsubstance was dissolved in 1 ml of ethanol, to which 200 μl of a 50%aqueous hydroxylamine solution was added. Following agitation at roomtemperature overnight, the solvent was distilled away, and the resultantwas purified using purification step A to obtain the title compound.

Yield: 7.49 mg

Example 39 Synthesis of5-[(4S)-4-amino-4-(hydroxycarbamoyl)butanamide]-3-chloro-2-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that5-amino-3-chloro-2-methylbenzenesulfonic acid was used instead of3-aminobenzenesulfonic acid in Example 38.

Yield: 5.4 mg

Example 40 Synthesis of3-[(4S)-4-amino-4-(hydroxycarbamoyl)butanamide]-5-chloro-4-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-amino-5-chloro-4-methylbenzenesulfonic acid was used instead of3-aminobenzenesulfonic acid in Example 38.

Yield: 7.88 mg

Example 41 Synthesis of3-[(4S)-4-amino-4-(hydroxycarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-amino-5-chloro-2-hydroxybenzenesulfonic acid was used instead of3-aminobenzenesulfonic acid in Example 38.

Yield: 5.7 mg

Example 42 Synthesis of(2S)-2-amino-4-[(6-sulfopyridine-2-yl)carbamoyl]butanoic acidtrifluoroacetic acid salt

50 mg (0.16 mmol) of Boc-Glu-OtBu, 35 mg (0.16 mmol) of1-(3-dimethylamino-propyl)-3-ethylcarbodiimide hydrochloride and 28 mg(0.16 mmol) of 1-hydroxybenzotriazole monohydrate were suspended in 1.0ml of methylene chloride, to which 28 mg (0.16 mmol) of6-amino-2-pyridinesulfonic acid was added. Following agitation at roomtemperature overnight, the resultant was subjected to extraction withethyl acetate/water. The organic layer was washed with saturated saline,and then added with sodium sulfate for drying. The organic layer wassubjected to vacuum condensation, 4.0 ml of TFA was added to theresulting residue, and the resultant was stirred overnight. 6.5 mg ofthe title compound was obtained using purification step A.

Yield: 6.5 mg

Example 43 Synthesis of(2S,3S)-2-amino-3-{[(3-sulfophenyl)carbamoyl]oxy}butanoic acid Step 1Synthesis of Boc-Allo-Thr-OMe

340 mg (2.0 mmol) of Allo-Thr-OMe hydrochloride was dissolved in 8.0 mlof methylene chloride, and added with 558 μl (4.0 mmol) of triethylamineand 436 mg (2.0 mmol) of di-tert-butyl dicarbonate. Following agitationat room temperature for 4 hours, the resultant was subjected to vacuumcondensation and extraction with ethyl acetate/water. The organic layerwas washed with saturated saline and then added with sodium sulfate fordrying. The organic layer was subjected to vacuum condensation to obtaina crude product of the title compound.

Yield: 460 mg

MS (ESI, m/z): 234 [M+H]+

Step 2 Synthesis of(2S,3S)-2-amino-3-{[(3-sulfophenyl)carbamoyl]oxy}butanoic acid

117 mg of the compound obtained in Step 1, 148 mg (0.5 mmol) oftriphosgene and 87 mg (0.5 mmol) of 3-aminobenzenesulfonic acid weredissolved in 2.0 ml of methylene chloride, added with 70 μl (0.5 mmol)of triethylamine and stirred at room temperature overnight. Followingaddition of 1.0 ml of ammonia-methanol solution, the solvent wasdistilled away. Purification step A was employed to obtain a crudeproduct of(2S,3S)-2-(N-tert-butoxycarbonylamino)-3-{[(3-sulfophenyl)carbamoyl]oxy}butanicacid methyl ester. To this, 20 mg (1.2 mmol) of lithium hydroxide and2.0 ml of water were added and stirred at room temperature overnight.Subsequently, 4.0 ml of TFA was added and stirred at room temperaturefor another 5 hours. After distilling the solvent away, purificationstep A was employed to obtain 5.2 mg of the title compound.

Yield: 5.2 mg

Example 44 Synthesis of(2S,3S)-2-amino-3-{[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]oxy}butanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 43 was replaced with3-amino-5-chloro-2-hydroxybenzenesulfonic acid.

Yield: 32.1 mg

Example 45 Synthesis of(2S,3R)-2-amino-3-{[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]oxy}butanoicacid

The title compound was obtained through a similar operation except thatBoc-Allo-Thr-OMe and 3-aminobenzenesulfonic acid were replaced withBoc-Thr-OMe and 3-amino-5-chloro-2-hydroxybenzenesulfonic acid,respectively, after Step 2 in Example 43.

Yield: 33.4 mg

Example 46 Synthesis of(2S)-2-amino-3-{[(3-sulfophenyl)carbamothioyl]amino}propanoic acid

35 mg (0.2 mmol) of 3-aminobenzenesulfonic acid and 57 mg (0.68 mmol) ofsodium hydrogen carbonate were added to a mixed solvent of 2.0 ml ofwater and 0.5 ml of THF, and stirred at room temperature for 15 minutes.20 μl (0.26 mmol) of thiophosgene was added and stirred at roomtemperature for another 40 minutes. Subsequently, 60 mg (0.2 mmol) oftert-butyl (2S)-3-amino-2-{[(tert-butoxy)carbonyl]amino}propanoatehydrochloride was added and stirred overnight. Following extraction withethyl acetate, the solvent was distilled away, and 4.0 ml oftrifluoroacetic acid was added to the residue, and the resultant wasstirred at room temperature for 5 hours. After distilling the solventaway, purification step A was used to obtain 3.5 mg of the titlecompound.

Yield: 3.5 mg

Example 47 Synthesis of(2S)-2-amino-3-{[(3-chloro-2-methyl-5-sulfophenyl)carbamothioyl]amino}propanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 46 was replaced with3-amino-5-chloro-4-methylbenzenesulfonic acid.

Yield: 6.9 mg

Example 48 Synthesis of(2S)-2-amino-3-{[(3-chloro-4-methyl-5-sulfophenyl)carbamothioyl]amino}propanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 46 was replaced with5-amino-3-chloro-2-methylbenzenesulfonic acid.

Yield: 5.8 mg

Example 49 Synthesis of3-[(4S)-4-amino-4-(carbamoyl)butanamide]benzene-1-sulfonic acid Step 1Synthesis of (4S)-4-(N-tert-butoxycarbonylamino)-4-(carbamoyl)butanoicacid

1010 mg (3.0 mmol) of Boc-Glu (OBzl)-OH, 316 μl (3.3 mmol) of ethylchloroformate and 460 μl (3.3 mmol) of triethylamine were suspended in12.0 ml of tetrahydrofuran, to which 2.0 ml of a concentrated aqueousammonia solution was added. Following agitation at room temperatureovernight, the resultant was subjected to extraction with ethylacetate/water. The organic layer was washed with saturated saline andthen sodium sulfate was added for drying. The organic layer wassubjected to vacuum condensation, and the resulting residue was addedwith 100 mg of 10% Pd/C and 12.0 ml of methanol and stirred at roomtemperature overnight in a hydrogen atmosphere under a normal pressure.After the reaction, the catalyst was filtrated away and the solvent wasdistilled away to obtain a crude purified substance of the titlecompound.

MS (ESI, m/z): 275 [M+H]+

Step 2 Synthesis of3-[(4S)-4-amino-4-(carbamoyl)butanamide]-benzene-1-sulfonic acid

100 mg (0.37 mmol) of the compound obtained in Step 1, 190 mg (0.5 mmol)of O-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate, 70 mg (0.5 mmol) of 1-hydroxy-7-azabenzotriazoleand 65 mg (0.37 mmol) of 3-aminobenzenesulfonic acid were suspended in2.0 ml of methylene chloride, added with 0.5 ml pyridine and stirred atroom temperature overnight. After distilling the solvent away,purification step A was used to obtain a crude product of3-[(4S)-4-(N-tert-butoxycarbonylamino)-4-(carbamoyl)butanamide]-benzene-1-sulfonicacid. To this, 4.0 ml of TFA was added and stirred at room temperaturefor 2 hours. After distilling the solvent away, purification step A wasused to obtain 1.6 mg of the title compound.

Yield: 1.6 mg

Example 50 Synthesis of3-[(4S)-4-amino-4-(carbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 49 was replaced with3-amino-5-chloro-2-hydroxybenzenesulfonic acid.

Yield: 4.1 mg

Example 51 Synthesis of5-[(4S)-4-amino-4-(carbamoyl)butanamide]-3-chloro-2-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 49 was replaced with3-amino-5-chloro-6-methylbenzenesulfonic acid.

Yield: 5.3 mg

Example 52 Synthesis of3-[(4S)-4-amino-4-(5-ethyl-1,3,4-oxadiazole-2-yl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid Step 1 Synthesis of(4S)-4-(N-tert-butoxycarbonylamino)-4-(N′-propionylcarbohydrazide)butanoic acid benzyl ester

674 mg (2.0 mmol) of Boc-Glu (OBzl)-OH, 420 mg (2.2 mmol) of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 336 mg (2.2mmol) of 1-hydroxybenzotriazole monohydrate and 230 mg (2.6 mmol) ofpropionic acid hydrazide were suspended in 5.0 ml of methylene chloride,to which 557 μl (4.0 mmol) of triethylamine was added. Followingagitation at room temperature overnight, vacuum condensation wasperformed and purification step A was used to obtain a crude purifiedsubstance of the title compound.

Yield: 500 mg

MS (ESI, m/z): 380 [M+H]+

Step 2 Synthesis of(4S)-4-(N-tert-butoxycarbonylamino)-4-(5-ethyl-1,3,4-oxadiazole-2-yl)butanoicacid

500 mg of the compound obtained in Step 1 and 182 mg (0.76 mmol) ofBurgess reagent were dissolved in 4.0 ml methylene chloride, and stirredat room temperature overnight. After distilling the solvent away, silicagel column chromatography was used for partial purification. To theresidue, 50 mg of 10% Pd/C and 4.0 ml of ethyl acetate were added andthe resultant was stirred at room temperature overnight in a hydrogenatmosphere under a normal pressure. The catalyst was filtrated awayafter the reaction, and the solvent was distilled away to obtain a crudepurified substance of the title compound.

Yield: 270 mg

MS (ESI, m/z): 300[M+H]+

Step 3 Synthesis of3-[(4S)-4-amino-4-(5-ethyl-1,3,4-oxadiazole-2-yl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

270 mg of the compound obtained in Step 2, 266 mg (0.7 mmol) ofO-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate, 100 mg (0.7 mmol) of 1-hydroxy-7-azabenzotriazoleand 160 mg (0.7 mmol) of 3-amino-5-chloro-2-hydroxybenzenesulfonic acidwere suspended in 4.0 ml of methylene chloride, to which 0.3 ml oftriethylamine was added and the resultant was stirred at roomtemperature overnight. After distilling the solvent away, purificationstep A was used to obtain a crude product of3-[(4S)-4-(N-tert-butoxycarbonylamino)-4-(5-ethyl-1,3,4-oxadiazole-2-yl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid. To this, 4.0 ml of TFA was added and stirred at room temperaturefor 2 hours. After distilling the solvent away, purification step A wasused to obtain 9.0 mg of the title compound.

Yield: 9.0 mg

Example 53 Synthesis of3-[(4S)-4-amino-4-(N′-propanoylhydrazinecarbonyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The compound was obtained as a by-product in Step 3 in Example 52.

Yield: 12.8 mg

Example 54 Synthesis of3-[(4S)-4-amino-4-(5-propyl-1,3,4-oxadiazole-2-yl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatpropionic acid hydrazide used in Step 1 in Example 52 was replaced withbutyric acid hydrazide.

Yield: 12.0 mg

Example 55 Synthesis of3-[(4S)-4-amino-4-[5-(propane-2-yl)-1,3,4-oxadiazole-2-yl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatpropionic acid hydrazide used in Step 1 in Example 52 was replaced withisobutyric acid hydrazide.

Yield: 11.8 mg

Example 56 Synthesis of3-[(4S)-4-amino-4-(ethylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid Step 1 Synthesis of(4S)-4-(N-tert-butoxycarbonylamino)-4-(ethylcarbamoyl)butanoic acid

337 mg (1.0 mmol) of Boc-Glu (OBzl)-OH, 191 mg (1.0 mmol) of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 152 mg (1.0mmol) of 1-hydroxybenzotriazole monohydrate and 0.5 ml of triethylaminewere suspended in 4.0 ml of DMF, to which 150 μl of 33% aqueousethylamine solution was added. Following agitation at room temperatureovernight, extraction was performed with ethyl acetate/water. Theorganic layer was washed with saturated saline and added with sodiumsulfate for drying. The organic layer was subjected to vacuumcondensation. The resulting residue was dissolved in 4.0 ml of 10%aqueous sodium hydroxide solution and stirred overnight. Purificationstep A was used to obtain a crude product of the title compound.

MS (ESI, m/z): 275 [M+H]+

Step 2 Synthesis of3-[(4S)-4-amino-4-(ethylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

220 mg (0.8 mmol) of the compound obtained in Step 1, 300 mg (0.8 mmol)of 0-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate, 110 mg (0.8 mmol) of 1-hydroxy-7-azabenzotriazoleand 180 mg (0.8 mmol) of 3-amino-5-chloro-2-hydroxybenzenesulfonic acidwere suspended in 4.0 ml of methylene chloride, to which 0.5 ml oftriethylamine was added and stirred at room temperature overnight. Afterdistilling the solvent away, purification step A was used to obtain 160mg of a crude product of3-[(4S)-4-(N-tert-butoxycarbonylamino)-4-(ethylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid. 144 mg of this crude product was dissolved in 4.0 ml of TFA andstirred at room temperature for 2 hours. After distilling the solventaway, purification step A was used to obtain 5.8 mg of the titlecompound.

Yield: 5.8 mg

Example 57 Synthesis of3-[(4S)-4-amino-4-(butylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatan aqueous ethylamine solution used in Step 1 in Example 56 was replacedwith n-butylamine.

Yield: 11.5 mg

Example 58 Synthesis of3-[(4S)-4-amino-5-(morpholine-4-yl)-5-oxopentanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatan aqueous ethylamine solution used in Step 1 in Example 56 was replacedwith morpholine.

Yield: 7.6 mg

Example 59 Synthesis of3-[(4S)-4-amino-4-(cyclohexylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatan aqueous ethylamine solution used in Step 1 in Example 56 was replacedwith cyclohexylamine.

Yield: 3.9 mg

Example 60 Synthesis of3-[(4S)4-amino-4-(cycloheptylcarbamoyl)butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except thatan aqueous ethylamine solution used in Step 1 in Example 56 was replacedwith cycloheptylamine.

Yield: 1.6 mg

Example 61 Synthesis of3-[(4S)-4-amino-4-[(benzenesulfonyl)carbamoyl]butanamide]benzene-1-sulfonicacid Step 1 Synthesis of(4S)-4-(N-tert-butoxycarbonylamino)-4-[(benzenesulfonyl)carbamoyl]butanoicacid

1.7 g (5.0 mmol) of Boc-Glu (OBzl)-OH, 1.03 g (5.0 mmol) ofdicyclohexylcarbodiimide and 611 mg (5.0 mmol) of4-(dimethylamino)pyridine were suspended in 20 ml of methylene chloride,to which 866 mg (5.0 mmol) of benzenesulfonamide was added. Followingagitation at room temperature overnight, extraction was performed withethyl acetate/1N hydrochloric acid. The organic layer was washed withsaturated saline and then added with sodium sulfate for drying. Theorganic layer was subjected to vacuum condensation. To the resultingresidue, 100 mg of 10% Pd/C and 20 ml of methanol were added and stirredovernight in a hydrogen atmosphere under a normal pressure. Afterfiltrating the catalyst away, the solvent was distilled away andpurification step A was used to obtain a crude product of the titlecompound.

MS (ESI, m/z): 387 [M+H]+

Step 2 Synthesis of3-[(4S)-4-amino-4-[(benzenesulfonyl)carbamoyl]butanamide]benzene-1-sulfonicacid

100 mg of the compound obtained in Step 1, 114 mg (0.3 mmol) ofO-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate, 40 mg (0.8 mmol) of 1-hydroxy-7-azabenzotriazoleand 55 mg (0.3 mmol) of 3-aminobenzenesulfonic acid were suspended in1.0 ml of methylene chloride, to which 0.1 ml of triethylamine was addedand stirred at room temperature overnight. After distilling the solventaway, purification step A was used to obtain a crude product of3-[(4S)-4-(N-tert-butoxycarbonylamino)-4-[(benzenesulfonyl)carbamoyl]butanamide]benzene-1-sulfonicacid. This crude product was dissolved in 4.0 ml of TFA and theresultant was stirred at room temperature for 2 hours. After distillingthe solvent away, purification step A was used to obtain 14.1 mg of thetitle compound.

Yield: 14.1 mg

Example 62 Synthesis of3-[(4S)-4-amino-4-[(benzenesulfonyl)carbamoyl]butanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Step 2 in Example 61 was replacedwith 3-amino-5-chloro-2-hydroxybenzenesulfonic acid.

Yield: 3.2 mg

Example 63 Synthesis of3-[(4S)-4-amino-4-[(benzenesulfonyl)carbamoyl]butanamide]-5-chloro-4-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Step 2 in Example 61 was replacedwith 3-amino-5-chloro-4-hydroxybenzenesulfonic acid.

Yield: 31.4 mg

Example 64 Synthesis of3-[(4S)-4-amino-4-(methylcarbamoyl)butanamide]benzene-1-sulfonic acidStep 1 Synthesis of(4S)-4-(N-tert-butoxycarbonylamino)-4-(methylcarbamoyl)butanoic acid

1010 mg (3 mmol) of Boc-Glu (OBzl)-OH, 600 mg (3.2 mmol) of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 460 mg(3.1 mmol) of 1-hydroxybenzotriazole monohydrate were suspended in 12 mlof DMF, to which 300 μl (2.0 mmol) of a 40% aqueous methylamine solutionwas added. Following agitation at room temperature overnight, extractionwas performed with ethyl acetate/water. The organic layer was washedwith saturated saline, and the resultant was added with sodium sulfatefor drying. The organic layer was subjected to vacuum condensation, andthe resultant residue was dissolved in 10 ml of methanol, added with 100mg of 10% Pd/C, and then stirred overnight in a hydrogen atmosphere.After the catalyst was filtrated away and the solvent was distilledaway, purification step A was used to obtain a crude product of thetitle compound.

MS (ESI, m/z): 261 [M+H]+

Step 2 Synthesis of3-[(4S)-4-amino-4-(methylcarbamoyl)butanamide]benzene-1-sulfonic acid

100 mg of the compound obtained in Step 1, 76 mg (0.2 mmol) ofO-(7-azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate, 28 mg (0.2 mmol) of 1-hydroxy-7-azabenzotriazoleand 36 mg (0.2 mmol) of 3-aminobenzenesulfonic acid were suspended in1.0 ml of methylene chloride, to which 0.1 ml of triethylamine was addedand the resultant was stirred at room temperature overnight. Afterdistilling the solvent away, purification step A was used to obtain acrude product of3-[(4S)-4-(N-tert-butoxycarbonylamino)-4-(methylcarbamoyl)butanamide]benzene-1-sulfonicacid. This crude product was dissolved in 4.0 ml of TFA, and stirred atroom temperature for 2 hours. After distilling the solvent away,purification step A was used to obtain 9.9 mg of the title compound.

Yield: 9.9 mg

Example 65 Synthesis of3-[(4S)-4-amino-4-(methylcarbamoyl)butanamide]-5-chloro-4-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Step 2 in Example 64 was replacedwith 3-amino-5-chloro-4-methylbenzenesulfonic acid.

Yield: 4.1 mg

Example 66 Synthesis of3-({[(2S)-2-amino-3-methoxy-3-oxopropyl]carbamoyl}amino)benzene-1-sulfonicacid

To 127 mg (0.5 mmol) of(2S)-3-amino-2-{[(tert-butoxy)carbonyl]amino}propanoic acid methylesterhydrochloride, 87 mg (0.5 mmol) of 3-aminobenzenesulfonic acid and 97 mg(0.6 mmol) of N,N-carbonyl diimidazole, 1 ml of methylene chloride and 1ml of tetrahydrofuran were added and stirred at room temperatureovernight. After distilling the solvent away, purification was carriedout using purification step A to obtain a crude purified substance ofthe title compound in protected form. To the obtained crude purifiedsubstance, 1 ml of methylene chloride and 1 ml of trifluoroacetic acidwere added and stirred at room temperature for 5 hours. After distillingthe solvent away, purification was carried out by purification step A toobtain the title compound.

Yield: 33.15 mg

Example 67 Synthesis of3-({[(2S)-2-amino-3-methoxy-3-oxopropyl]carbamoyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 66 was replaced with3-amino-5-chloro-2-hydroxybenzenesulfonic acid.

Yield: 19.1 mg

Example 68 Synthesis of5-({[(2S)-2-amino-3-methoxy-3-oxopropyl]carbamoyl}amino)-3-chloro-2-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 66 was replaced with5-amino-3-chloro-2-methylbenzenesulfonic acid.

Yield: 11.84 mg

Example 69 Synthesis of3-({[(2S)-2-amino-3-methoxy-3-oxopropyl]carbamoyl}amino)-5-chloro-4-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 66 was replaced with3-amino-5-chloro-4-methylbenzenesulfonic acid.

Yield: 14.57 mg

Example 70 Synthesis of 3-({[(2S)-2-amino2-(methylcarbamoyl)ethoxy]carbonyl}amino)benzene-1-sulfonic acid Step 1Synthesis of t-butylN-[(1S)-2-hydroxy-1-(methylcarbamoyl)ethyl]carbamate

885 mg (3.0 mmol) of Boc-Ser (OBzl)-OH, 600 mg (3.2 mmol) of1-(3-dimethylamino-propyl)-3-ethylcarbodiimide hydrochloride and 460 mg(3.1 mmol) of 1-hydroxybenzotriazole monohydrate were dissolved in 12 mlof DMF, to which 300 μl of 40% aqueous methylamine solution was added.Following agitation at room temperature overnight, extraction wasperformed with ethyl acetate/1N hydrochloric acid. After washing theorganic layer with saturated saline, sodium sulfate was added fordrying. The organic layer was subjected to vacuum condensation, and theresulting residue was dissolved in 12 ml of methanol, to which 100 mg of10% Pd/C was added and stirred overnight in a hydrogen atmosphere.Purification step A was used to obtain a crude product of the titlecompound.

MS (ESI, m/z): 219 [M+H]+

Step 2 Synthesis of3-({[(2S)-2-amino-2-(methylcarbamoyl)ethoxy]carbonyl}amino)benzene-1-sulfonicacid

80 mg (0.37 mmol) of the compound obtained in Step 1, 40 mg (0.1 mmol)of triphosgene and 60 mg (0.35 mmol) of 3-aminobenzenesulfonic acid weresuspended in 2.0 ml of methylene chloride, to which 0.5 ml of pyridinewas added and stirred at room temperature overnight. After distillingthe solvent away, purification step A was used to obtain anintermediate. This crude product was dissolved in 4.0 ml oftrifluoroacetic acid and stirred at room temperature for 15 minutes.After distilling the solvent away, purification step A was used toobtain 6.79 mg of the title compound.

Yield: 6.79 mg

Example 71 Synthesis of3-({[(2S)-2-amino-2-(methylcarbamoyl)ethoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 70 was replaced with3-amino-5-chloro-2-hydroxybenzenesulfonic acid.

Yield: 26.3 mg

Example 72 Synthesis of3-({[(2S)-2-amino-2-(methylcarbamoyl)ethoxy]carbonyl}amino)-5-chloro-4-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 70 was replaced with3-amino-5-chloro-4-methylbenzenesulfonic acid.

Yield: 0.8 mg

Example 73 Synthesis of3-({[(2S)-2-amino-2-[(benzenesulfonyl)carbamoyl]ethoxy]carbonyl}amino)benzene-1-sulfonicacid Step 1 Synthesis of t-butylN-[(1S)-1-[(benzenesulfonyl)carbamoyl]-2-hydroxyethyl]carbamate

885 mg (3.0 mmol) of Boc-Ser (OBzl)-OH, 620 mg (3.0 mmol) ofdicyclohexylcarbodiimide and 370 mg (3.0 mmol) of4-(dimethylamino)pyridine were dissolved in 12 ml of methylene chloride,to which 470 mg (3.0 mmol) of benzenesulfonamide was added. Followingagitation at room temperature overnight, extraction was performed withethyl acetate/1N hydrochloric acid. After washing the organic layer withsaturated saline, sodium sulfate was added for drying. The organic layerwas subjected to vacuum condensation. The resulting residue wasdissolved in 12 ml of methanol, added with 100 mg of 10% Pd/C andstirred overnight in a hydrogen atmosphere. Purification step A was usedto obtain a crude product of the title compound.

ESI (m/z): 345 [M+H]+

Step 2 Synthesis of3-({[(2S)-2-amino-2-[(benzenesulfonyl)carbamoyl]ethoxy]carbonyl}amino)benzene-1-sulfonicacid

80 mg (0.23 mmol) of the compound obtained in Step 1, 40 mg (0.1 mmol)of triphosgene and 40 mg (0.23 mmol) of 3-aminobenzenesulfonic acid weresuspended in 2.0 ml of methylene chloride, to which 0.5 ml of pyridinewas added and stirred at room temperature overnight. After distillingthe solvent away, purification step A was used to obtain anintermediate. This crude product was dissolved in 4.0 ml oftrifluoroacetic acid and stirred at room temperature for an hour. Afterdistilling the solvent away, purification step A was used to obtain 21.2mg of the title compound.

Yield: 21.2 mg

Example 74 Synthesis of3-({[(2S)-2-amino-2-[(benzenesulfonyl)carbamoyl]ethoxy]carbonyl}amino)-5-chloro-2-hydroxybenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 73 was replaced with3-amino-5-chloro-2-hydroxybenzenesulfonic acid.

Yield: 8.36 mg

Example 75 Synthesis of3-({[(2S)-2-amino-2-[(benzenesulfonyl)carbamoyl]ethoxy]carbonyl}amino)-5-chloro-4-methylbenzene-1-sulfonicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 73 was replaced with3-amino-5-chloro-4-methylbenzenesulfonic acid.

Yield: 8.8 mg

Example 76 Synthesis of(2S)-4-[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]-2-(dimethylamino)butanoicacid Step 1 Synthesis of(2S)-2-amino-4-[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]butanoicacid

101 mg (0.33 mmol) of Boc-Glu-OtBu, 130 mg (0.33 mmol) of HATU, 45 mg(0.33 mmol) of HOAt and 77 mg (0.33 mmol) of3-amino-5-chloro-2-hydroxybenzenesulfonic acid were suspended in 2.0 mlof DMF, to which 0.5 ml of pyridine was added and stirred at roomtemperature overnight. After distilling the solvent away, purificationstep A was used to obtain an intermediate. This crude product wasdissolved in 2.0 ml of TFA and stirred at room temperature for 90minutes. After distilling the solvent away, purification step A was usedto obtain the title compound.

Yield: 90 mg

ESI (m/z): 353, 355 [M+H]+

Step 2 Synthesis of(2S)-4-[(5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]-2-(dimethylamino)butanoicacid

35 mg (0.1 mmol) of the compound obtained in Step 1 and 30 mg of Pd/Cwere suspended in 1.0 ml of a 37% aqueous formaldehyde solution andstirred overnight in a hydrogen atmosphere. After filtrating Pd/C away,purification step A was used to obtain 11.2 mg of the title compound.

Yield: 11.2 mg

Example 77 Synthesis of(2S)-2-amino-3-{[(3-sulfophenyl)carbamoyl]amino}propanoic acid

100 mg of (2S)-3-amino-2-{[(tert-butoxy)carbonyl]amino}propanoic acidtert-butylester hydrochloride (Boc-DAP-OtBu.HCl) and 65 mg ofN,N′-carbonyl diimidazole (CDI) were dissolved in acetonitrile andstirred at room temperature for 10 minutes. To this, 60 mg of3-aminobenzenesulfonic acid was added and stirred at 50° C. for 5 hours.The solvent was distilled away, and the resultant was subjected toextraction with ethyl acetate to collect and dry the organic layer. Thesolvent was distilled away to obtain a crude product of the titlecompound in protected form. The obtained crude product was added with 5ml of trifluoroacetic acid and stirred overnight. The solvent wasdistilled away and the resulting residue was purified by purificationstep A to obtain the title compound.

Yield: 4.93 mg

Example 78 Synthesis of(2S)-4-[5-chloro-2-hydroxy-3-sulfophenyl)carbamoyl]-2-(methylamino)butanoicacid

To 75 mg of3-[(4S)-4-amino-5-(benzyloxy)-5-oxopentanamide]-5-chloro-2-hydroxybenzene-1-sulfonicacid, 1 ml of methylene chloride, 41 mg of 2-nitrobenzenesulfonylchloride and 50 μl of triethylamine were added. After agitation for anhour, extraction was performed with ethyl acetate to obtain 100 mg of acrude product. The obtained crude product was added with 23 mg ofpotassium carbonate, 1 ml of DMF and 10.5 μl of methyl iodide andstirred at room temperature overnight. The resultant was diluted withwater and acetonitrile, and subjected to partial purification bypurification step A. The resulting crude purified substance wasdissolved in 1 ml of THF, 0.5 ml of ethanol and 0.5 ml of water, towhich 10 mg of lithium hydroxide was added. While confirming thereaction progress, sodium hydroxide was appropriately added. At the endof the reaction, 2 ml of ethyl acetate was added and stirred. Thesolvent was distilled away, and the resultant was added with water andlyophilized. The resulting lyophilized product was dissolved in 1 ml ofDMF, to which 45 μl of 1-dodecanethiol and 60 μl of an ethanol solutionof 28% sodium ethoxide were added and the resultant was stirred for anhour. 45 μl of 1-dodecanethiol and 60 μl of an ethanol solution of 28%sodium ethoxide were further added and stirred. The reaction solutionwas diluted with water and acetonitrile, and then purified usingpurification step A to obtain the title compound.

Yield: 2.4 mg

Example 79 Synthesis of(2S)-3-{[(3-chloro-4-methyl-5-sulfophenyl)carbamoyl]amino}-2-(methylamino)propanoicacid

100 mg of Boc-DAP-OtBu hydrochloride and 60 mg of CDI were dissolved in1 ml of acetonitrile and stirred for 5 minutes. 75 mg of5-amino-3-chloro-2-methylbenzenesulfonic acid was added and stirredovernight. Extraction was performed with ethyl acetate to obtain a crudeproduct. The obtained crude product was added with 1.5 ml of dioxane and0.5 ml of dioxane solution containing 4N hydrochloric acid and stirredfor 2 hours. The solvent was distilled away to obtain a crude product.To the obtained crude product, 2 ml of methylene chloride was added, and52 mg of 2-nitrobenzenesulfonyl chloride and 0.14 ml of triethylaminewere added. After 2 hours of agitation, extraction was performed withethyl acetate to obtain a crude product. The obtained crude product wasadded with 50 mg of potassium carbonate, 2 ml of DMF and 0.1 ml ofmethyl iodide and stirred at room temperature overnight. Followingextraction with ethyl acetate, the solvent was distilled away to obtaina crude product. The obtained crude product was added with 3 ml oftrifluoroacetic acid and stirred at room temperature for 5 hours. Thesolvent was distilled away and the residue was lyophilized. Theresulting lyophilized product was added with 1 ml of DMF, further addedwith 54 μl of 1-dodecanethiol and 80 μl of a 28% sodium ethoxide ethanolsolution, and stirred at room temperature. Following dilution with waterand acetonitrile, purification was performed using purification step Ato obtain the title compound.

Yield: 18.4 mg

Example 80 Synthesis of(2S)-2-amino-3-{[(4-methyl-3-sulfophenyl)carbamoyl]amino}propanoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2-methylbenzenesulfonic acid.

Yield: 6.86 mg

Example 81 Synthesis of(2S)-2-amino-3-{[(4-methoxy-3-sulfophenyl)carbamoyl]amino}propanoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2-methoxybenzenesulfonic acid.

Yield: 3.5 mg

Example 82 Synthesis of(2S)-2-amino-3-{[(2-methoxy-5-sulfophenyl)carbamoyl]amino}propanoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-methoxybenzenesulfonic acid.

Yield: 6.22 mg

Example 83 Synthesis of(2S)-2-amino-3-{[(3-acetamide-2-hydroxy-5-sulfophenyl)carbamoyl]amino}propanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-(acetylamino)-5-amino-4-hydroxybenzenesulfonic acid.

Yield: 5.81 mg

Example 84 Synthesis of(2S)-2-amino-3-{[(2-hydroxy-3-nitro-5-sulfophenyl)carbamoyl]amino}propanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-hydroxy-5-nitrobenzenesulfonic acid.

Yield: 6.1 mg

Example 85 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)benzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-aminobenzoic acid.

Yield: 2.66 mg

Example 86 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}-amino)-2,5-dichlorobenzoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-2,5-dichlorobenzoic acid.

Yield: 7.56 mg

Example 87 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-methylbenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-2-methylbenzoic acid.

Yield: 10.76 mg

Example 88 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-4-chlorobenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-chlorobenzoic acid.

Yield: 2.3 mg

Example 89 Synthesis of(2S)-2-amino-3-{[(2,4-dimethyl-5-sulfophenyl)carbamoyl]amino}propanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2,4-dimethylbenzoic acid.

Yield: 1 mg

Example 90 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-hydroxybenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-aminosalicylic acid.

Yield: 10.5 mg

Example 91 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-4-methoxybenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-methoxybenzoic acid.

Yield: 4.72 mg

Example 92 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-5-nitrobenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-5-nitrobenzoic acid.

Yield: 3.5 mg

Example 93 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-4-methylbenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-methylbenzoic acid.

Yield: 20.4 mg

Example 94 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-4-hydroxybenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-hydroxybenzoic acid.

Yield: 5.6 mg

Example 95 Synthesis of5-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-chlorobenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2-chlorobenzoic acid.

Yield: 24.4 mg

Example 96 Synthesis of5-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-hydroxybenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2-hydroxybenzoic acid.

Yield: 2.75 mg

Example 97 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-4-fluorobenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-fluorobenzoic acid.

Yield: 31.92 mg

Example 98 Synthesis of5-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-methoxybenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2-methoxybenzoic acid.

Yield: 9.1 mg

Example 99 Synthesis of5-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-methylbenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2-methylbenzoic acid.

Yield: 4.3 mg

Example 100 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-methoxybenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-2-methoxybenzoic acid.

Yield: 2.35 mg

Example 101 Synthesis of3-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-5-methoxybenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-5-methoxybenzoic acid.

Yield: 2.42 mg

Example 102 Synthesis of5-({[(2S)-2-amino-2-carboxyethyl]carbamoyl}amino)-2-fluorobenzoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2-fluorobenzoic acid.

Yield: 16.73 mg

Example 103 Synthesis of(2S)-2-amino-3-{[(3,4-dimethyl-5-sulfophenyl)carbamoyl]amino}propanoicacid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with5-amino-2,3-dimethylbenzenesulfonic acid.

Yield: 3.33 mg

Example 104 Synthesis of(2S)-2-amino-3-{[(2-fluoro-5-sulfophenyl)carbamoyl]amino}propanoic acid

The title compound was obtained through a similar operation except that3-aminobenzenesulfonic acid used in Example 77 was replaced with3-amino-4-fluorobenzenesulfonic acid.

Yield: 2.54 mg

TABLE 1 Example MS Number Structure 1H-NMR (ESI, m/z) 1

224[M + H]⁺ 2

1H-NMR(300 MHz, D₂O) δ: 8.12(d, 1H), 7.3(d, 1H), 3.87(t, 1H),2.63-2.69(m, 2H), 2.55(s, 3H), 2.10-2.20(m, 2H) 317[M + H]⁺ 3

339[M + H]⁺ 4

1H-NMR(300 MHz, D₂O) δ: 7.68-7.80(m, 1H), 7.30-7.50(m, 3H), 4.00-4.19(m,1H), 2.97-3.68(m, 2H) 321[M + H]⁺ 5

1H-NMR(300 MHz, D₂O) δ: 10.98(s, 1H), 10.02(s, 1H), 8.30(br, 2H),7.70(s, 1H), 7.21(s, 1H), 4.05-4.25(m, 1H), 3.00-3.80(m, 2H) 371[M + H]⁺6

1H-NMR(300 MHz, D₂O) δ: 7.62(d, 1H), 7.34(d, 1H), 4.02(dd, 1H), 3.74(dd,1H), 3.59(dd, 1H) 354[M + H]+ 352[M − H]⁻ 7

1H-NMR(300 MHz, DMSO-d6) δ: 7.83(d, 1H,), 7.12(d, 1H), 4.17-4.22(m, 1H),4.02- 4.06(m, 1H), 1.19(d, 3H) 323[M − H]⁻ 8

1H-NMR(300 MHz, DMSO-d6) δ: 7.85(d, 1H), 7.12(d, 1H), 4.34(dd, 1H),4.11(dd, 1H), 1.90-2.00(m, 1H), 0.95-1.00(m, 6H) 353[M + H]+ 351[M − H]⁻9

1H-NMR(300 MHz, DMSO-d6) δ: 7.83(d, 1H), 7.29-7.40(m, 5H), 7.12(d, 1H),4.21(dd, 1H), 3.96(dd, 1H), 3.64-3.73(m, 1H), 2.89-2.94(m, 2H) 401[M +H]+ 399[M − H]⁻

TABLE 2 Example MS Number Structure 1H-NMR (ESI, m/z) 10

1H-NMR(300 MHz, DMSO-d6) δ: 7.80- 7.85(m, 1H), 7.46-7.52(m, 5H), 7.13(d,1H), 4.60-4.80(m, 1H), 4.30-4.44(m, 2H) 387[M + H]+ 385[M − H]⁻ 11

1H-NMR(300 MHz, D₂O) δ: 7.63(brs, 1H), 7.38(d, 1H), 4.35(dd, 1H),4.19(dd, 1H), 3.48-3.57(m, 1H), 2.33-2.39(m, 2H), 1.86- 1.94(m, 2H)382[M + H]+ 380[M − H]⁻ 12

1H-NMR(300 MHz, D₂O) δ: 7.68(brs, 1H), 7.39(dd, 1H), 4.41(dd, 1H)4.24(dd, 1H), 3.55-3.65(m, 1H), 2.61(t, 2H), 1.90-2.10(m, 2H) 364[M +H]+ 362[M − H]⁻ 13

1H-NMR(300 MHz, D₂O) δ: 7.62(d, 1H), 7.51(d, 1H), 3.88(dd, 1H), 3.71(dd,1H), 3.52(dd, 1H), 2.45(s, 3H) 352[M + H]+ 350[M − H]⁻ 14

1H-NMR(300 MHz, D₂O) δ: 7.60(d, 1H), 7.55(d, 1H), 3.82(dd, 1H), 3.69(d,1H), 3.51(dd, 1H), 2.20(s, 3H) 352[M + H]+ 350[M − H]⁻ 15

391[M + H]⁺ 16

1H-NMR(300 MHz, D₂O) δ: 7.52(d, 1H), 7.42(d, 1H), 7.17-7.30(m, 5H),4.60-4.80(m, 1H), 4.14(s, 2H), 2.55-2.60(m, 2H), 2.33- 2.40(m, 2H)467[M + H]+ 465[M − H]⁻ 17

1H-NMR(300 MHz, D₂O) δ: 7.66(d, 1H), 7.44(d, 1H), 7.20-7.31(m, 5H),3.98(t, 1H), 3.58(s, 2H), 2.6(t, 2H), 2.10-2.20(m, 2H) 485[M + H]+ 483[M− H]⁻

TABLE 3 Example MS Number Structure 1H-NMR (ESI, m/z) 18

1H-NMR(300 MHz, D₂O) δ: 7.66(s, 1H), 7.46-7.55(m, 1H), 7.35-7.40(m, 2H),7.10- 7.30(m, 5H), 4.78-4.83(m, 1H), 4.12(s, 2H), 2.47-2.53(m, 2H),2.32-2.39(m, 2H) 417[M + H]+ 415[M − H]⁻ 19

1H-NMR(300 MHz, D₂O) δ: 7.15-7.62(m, 6H), 4.70-4.80(m, 1H), 2.60-2.73(m,2H), 2.40-2.52(m, 2H) 533[M + H]⁺ 20

1H-NMR(300 MHz, D₂O) δ: 7.17-7.60(m, 8H), 4.77-4.90(m, 1H), 2.55(m, 2H),2.40- 2.47(m, 2H) 481, 483 [M + H]⁺ 21

1H-NMR(300 MHz, DMSO-d6) δ: 11.07(brs, 1H), 9.39(s, 1H), 8.01-8.05(m,2H), 7.96(d, 1H), 7.59-7.69(m, 3H), 7.12(d, 1H), 4.89(t, 1H),2.60-2.73(m, 2H), 2.27-2.37(m, 2H) 453, 455[M + H]+ 451, 453[M − H]⁻ 22

1H-NMR(300 MHz, DMSO-d6) δ: 11.10(s, 1H), 9.37(s, 1H), 8.91(t, 1H),8.16(m, 2H), 8.03(m, 1H), 7.23-7.40(m, 6H), 7.15(d, 1H), 4.20-4.50(m,2H), 3.80-3.85(m, 1H), 2.50- 2.60(m, 2H), 2.05(dd, 2H) 442, 444[M + H]+440[M − H]⁻ 23

1H-NMR(300 MHz, DMSO-d6) δ: 11.11(s, 1H), 10.31(s, 1H), 8.45-8.54(m,1H), 8.00- 8.20(m, 3H), 7.12-7.33(m, 6H), 3.70- 3.80(m, 1H),3.40-3.50(m, 2H), 2.71- 2.80(m, 2H), 2.40-2.60(m, 2H), 1.90- 2.08(m, 2H)456, 458[M + H]+ 454, 456[M − H]⁻

TABLE 4 Example MS Number Structure 1H-NMR (ESI, m/z) 24

1H-NMR(300 MHz, DMSO-d6) δ: 9.38(s, 1H), 8.40(m, 1H), 8.03(m, 1H),7.10-7.31(m, 7H), 3.68-3.78(m, 1H), 3.10-3.20(m, 2H), 2.40-2.70(m, 4H),1.85-2.06(m, 2H), 1.70- 1.80(m, 2H) 470, 472[M + H]+ 468, 471[M − H]⁻ 25

1H-NMR(300 MHz, DMSO-d6) δ: 9.36(s, 1H), 8.30-8.40(m, 1H), 8.00-8.05(m,1H), 7.10-7.30(m, 7H), 3.70(t, 1H), 3.30-3.45(m, 2H), 3.10-3.20(m, 2H),2.50-2.65(m, 2H), 1.90-2.05(m, 2H), 1.40-1.65(m, 4H) 484, 486[M + H]+482, 484[M − H]⁻ 26

1H-NMR(300 MHz, DMSO-d6) δ: 11.07(brs, 1H), 9.41(s, 1H), 8.80(m, 3H),7.98(d, 1H), 7.79(d, 2H), 7.12(d, 1H), 6.83(d, 2H), 4.87(br, 1H),3.02(s, 6H), 2.55-2.75(m, 2H), 2.27-2.36(m, 2H) 496, 498[M + H]+ 494[M −H]⁻ 27

1H-NMR(300 MHz, DMSO-d6) δ: 11.07(br, 1H), 9.38(s, 1H), 7.99(m, 2H),7.83(dd, 1H), 7.31(dd, 1H), 7.12(d, 1H), 4.80(m, 1H), 2.20-2.80(m, 4H)459[M + H]+ 457[M − H]⁻ 28

318[M − H]⁻ 29

308[M − H]⁻

TABLE 5 Example MS Number Structure 1H-NMR (ESI, m/z) 30

1H-NMR(300 MHz, DMSO-d6) δ: 11.09(s, 1H), 10.83(s, 1H), 9.36(s, 1H),8.53(t, 1H), 8.00-8.30(m, 4H), 6.96-7.60(m, 5H), 3.76- 3.79(m, 1H),3.20-3.50(m, 2H), 2.85- 2.92(m, 2H), 2.40-2.60(m, 2H), 1.97- 2.07(m, 2H)495[M + H]⁺ 31

1H-NMR(300 MHz, DMSO-d6) δ: 9.38(s, 1H), 8.47(t, 1H), 7.90-8.20(m, 3H),7.14- 7.23(m, 2H), 6.70-6.81(m, 2H), 3.70- 3.80(m, 3H), 3.40-3.50(m,2H), 2.74(t, 2H), 2.40-2.55(m, 2H), 1.90-2.01(m, 2H) 488[M + H]⁺ 32

490, 492[M + H]+ 488[M − H]⁻ 33

1H-NMR(400 MHz, D₂O) δ: 8.57(s, 1H), 7.73(d, 1H), 7.41(d, 1H),7.32-7.37(m, 2H), 4.11(t, 2H), 3.96(dd, 1H), 3.07-3.25(m, 2H), 2.63(t,2H), 2.15-2.30(m, 2H), 1.89-2.00(m, 2H) 460, 461[M + H]+ 458[M − H]⁻ 34

1H-NMR(300 MHz, D₂O) δ: 7.66(d, 1H), 7.42(d, 1H), 3.8(t, 1H), 2.99(s,3H), 2.54- 2.62(m, 2H), 2.08-2.20(m, 2H) 430, 432[M + H]+ 428, 430[M −H]⁻ 35

1H-NMR(300 MHz, D₂O) δ: 7.76-7.77(m, 1H), 7.20-7.51(m, 4H), 3.99(t, 1H),3.16(s, 3H), 2.54-2.60(m, 2H), 2.16-2.24(m, 2H) 380[M + H]⁺

TABLE 6 Example MS Number Structure 1H-NMR (ESI, m/z) 36

1H-NMR(300 MHz, D₂O) δ: 7.72(d, 1H), 7.53(d, 1H), 4.01(t, 1H), 3.19(s,3H), 2.50- 2.56(m, 2H), 2.46(s, 3H), 2.14-2.23(m, 2H) 428[M + H]+ 426[M− H]⁻ 37

1H-NMR(300 MHz, D₂O) δ: 7.66(d, 1H), 7.49(d, 1H), 3.99(t, 1H), 3.16(s,3H), 2.63(t, 2H), 2.15-2.30(m, 5H) 428[M + H]+ 426[M − H]⁻ 38

1H-NMR(400 MHz, D₂O) δ 7.35-7.75(m, 4H), 3.82(t, 1H), 2.38-2.56(m, 2H),2.05- 2.15(m, 2H) 318[M + H]⁺ 39

1H-NMR(400 MHz, D₂O) δ: 7.70(s, 1H), 7.62(s, 1H), 3.70-3.83(m, 1H),2.35- 2.50(m, 5H), 2.02-2.15(m, 2H) 366[M + H]+ 364[M − H]⁻ 40

1H-NMR(400 MHz, D₂O) δ: 7.64(d, 1H), 7.45(d, 1H), 3.74(t, 1H),2.45-2.55(m, 2H), 2.14(s, 3H), 2.04-2.10(m, 2H) 366[M + H]⁺ 41

1H-NMR(400 MHz, D₂O) δ: 7.65(d, 1H), 7.42(d, 1H), 3.75-3.83(m, 1H),2.40- 2.60(m, 2H), 2.04-2.14(m, 2H) 368[M + H]+ 366[M − H]⁻ 42

303[M + H]⁺

TABLE 7 Example MS Number Structure 1H-NMR (ESI, m/z) 43

1H-NMR(300 MHz, DMSO-d6): δ 7.58- 7.12(m, 4H), 4.20-4.03(m, 2H), 1.08(m, 3H) 319[M + H]⁺ 44

1H-NMR(300 MHz, DMSO-d6): δ 8.08 (s, 1H), 6.97(s, 1H), 4.22-4.06(m, 2H),1.08(m, 3H) 369, 371[M + H]⁺ 45

1H-NMR(400 MHz, DMSO-d6): δ 8.08 (s, 1H), 6.97(s, 1H), 4.22-4.06(m, 2H),1.08(m, 3H) 369, 371[M + H]⁺ 46

1H-NMR(300 MHz, D₂O): δ 7.65-7.41(m, 4H), 4.21-4.16(m, 2H), 4.01-3.95(m,1H) 320[M + H]⁺ 47

1H-NMR(400 MHz, DMSO-d6): δ 9.62(s, 1H), 8.24(s, 3H), 7.67(s, 1H),7.49(s, 1H), 7.38(s, 1H), 4.21-4.03(m, 1H), 4.03-4.00(m, 1H),3.86-3.79(m, 1H), 2.18(s, 3H) 368, 370[M + H]⁺ 48

1H-NMR(400 MHz, DMSO-d6): δ 9.98(s, 1H), 8.26(s, 3H), 7.81(s, 1H),7.77(s, 1H), 7.62(s, 1H), 4.22-4.15(m, 1H), 4.10-4.07(m, 1H),3.84-3.78(m, 1H), 2.53(s, 3H) 368, 370[M + H]⁺ 49

302[M + H]⁺

TABLE 8 Example MS Number Structure 1H-NMR (ESI, m/z) 50

1H-NMR(400 MHz, DMSO-d6): δ 11.1(s, 1H), 9.41(s, 1H), 8.05(s, 3H),8.01(s, 1H) 7.85(s, 1H), 7.62(s, 1H), 7.15(s, 1H), 3.77-3.75(m, 1H),2.55-2.53(m, 2H), 2.06-1.99(m, 2H) 352, 354[M + H]⁺ 51

1H-NMR(400 MHz, DMSO-d6): δ 10.19 (s, 1H), 8.08(s, 3H), 7.94(s, 1H),7.87 (s, 1H) 7.80(s, 1H), 7.64(s, 1H), 3.79- 3.76(m, 1H), 2.50(s, 3H),2.46-2.40(m, 2H), 2.06-1.99(m, 2H) 350, 352[M + H]⁺ 52

1H-NMR(400 MHz, DMSO-d6): δ 11.6(s, 1H), 11.2(s, 1H), 10.6(s, 1H),10.5(s, 1H) 9.93(s, 1H), 8.01(s, 1H), 7.20(s, 1H), 4.97-4.94(m, 1H),3.02- 2.99(m, 2H), 2.58-2.50(m, 2H), 2.25- 2.20(m, 2H), 1.07-1.04(m, 3H)405, 407[M + H]⁺ 53

1H-NMR(400 MHz, DMSO-d6): δ 11.2(s, 1H), 10.1(s, 1H), 9.83(s, 1H),9.71(s, 1H) 8.25(s, 3H), 8.00(s, 1H), 7.22(s, 1H), 4.25-4.21(m, 1H),2.32- 2.28(m, 2H), 2.15-2.03(m, 4H), 1.03- 0.99(m, 3H) 423, 425[M + H]⁺54

419[M + H]⁺ 55

419[M + H]⁺ 56

1H-NMR(300 MHz, D₂O): δ 7.69(s, 1H), 7.44(s, 1H), 3.91-3.84(m, 1H),3.15-3.08(m, 2H), 2.56-2.52(m, 2H), 2.15-2.09(m, 2H), 1.01-0.96(m, 3H)380, 382[M + H]⁺

TABLE 9 Example MS Number Structure 1H-NMR (ESI, m/z) 57

1H-NMR(300 MHz, D₂O): δ 7.72(s, 1H), 7.43(s, 1H), 3.91-3.84(m, 1H),3.12- 3.03(m, 2H), 2.58-2.52(m, 2H), 2.15- 2.09(m, 2H), 1.37-1.26(m,2H), 1.25- 1.09(m, 2H), 0.74-0.68(m, 3H) 408, 410[M + H]⁺ 58

1H-NMR(300 MHz, D₂O): δ 7.66(s, 1H), 7.43(s, 1H), 4.53-4.47(m, 1H),3.66- 3.49(m, 8H), 2.61-2.57(m, 2H), 2.15- 2.11(m, 2H) 422, 424[M + H]⁺59

1H-NMR(400 MHz, DMSO-d6): δ 11.01 (s, 1H), 9.39(s, 1H), 8.29(s, 1H),8.08 (s, 3H), 8.01(s, 1H), 7.14(s, 1H), 3.73-3.70(m, 1H), 3.59-3.53(m,2H), 2.03-1.96(m, 2H), 1.81-1.69(m, 5H), 1.29-1.16(m, 6H) 434, 436[M +H]⁺ 60

1H-NMR(300 MHz, CD₃OD): δ 8.08(s, 1H), 7.26(s, 1H), 3.77-3.76(m, 1H),3.70-3.67(m, 1H), 2.54-2.53(m, 2H), 2.07-2.02(m, 2H), 1.59-1.38(m, 12H)448, 450[M + H]⁺ 61

1H-NMR(300 MHz, DMSO-d6): δ 8.18- 7.21(m, 9H), 3.89-3.81(m, 1H), 2.41-2.33(m, 2H), 2.04-1.96(m, 2H) 442[M + H]⁺ 62

1H-NMR(300 MHz, CD₃OD): δ 8.18-7.37 (m, 7H), 3.95-3.87(m, 1H), 2.61-2.48(m, 2H), 2.20-2.08(m, 2H) 492, 494[M + H]⁺

TABLE 10 Example MS Number Structure 1H-NMR (ESI, m/z) 63

1H-NMR(300 MHz, DMSO-d6): δ 8.18- 7.39(m, 7H), 3.89-3.81(m, 1H), 2.41-2.36(m, 2H), 2.16(s, 3H), 2.04-1.96(m, 2H) 490, 492[M + H]⁺ 64

1H-NMR(300 MHz, CD₃OD): δ 8.04(s, 1H), 7.67-7.33(m, 3H), 3.95-3.91(m,1H), 2.81(s, 3H), 2.59-2.49(m, 2H), 2.19-2.13(m, 2H) 316[M + H]⁺ 65

1H-NMR(300 MHz, CD₃OD): δ 7.74(s, 1H), 7.70(s, 1H), 3.95-3.91(m, 1H),2.82(s, 3H), 2.65-2.59(m, 2H), 2.28(s, 3H), 2.21-2.15(m, 2H) 364,366[M + H]⁺ 66

1H-NMR(400 MHz, D₂O) δ: 7.66(m, 1H), 7.35-7.48(m, 3H), 4.24(t, 1H),3.79(s, 3H), 3.70-3.74(m, 2H) 318[M + H]+ 316[M − H]⁻ 67

1H-NMR(400 MHz, D₂O) δ: 7.63-7.66(m, 1H), 7.38-7.41(m, 1H), 4.20-4.29(m,1H), 3.50-3.82(m, 5H) 368, 370[M + H]+ 366[M − H]⁻ 68

1H-NMR(400 MHz, D₂O) δ: 7.66(d, 1H), 7.53(d, 1H), 4.25(t, 1H), 3.80(s,3H), 3.66- 3.79(m, 2H), 2.52(s, 3H) 366, 368[M + H]⁺ 69

1H-NMR(400 MHz, D₂O) δ: 7.67(d, 1H), 7.57(d, 1H), 4.25(t, 1H), 3.80(s,3H), 3.72(d*2, 2H), 2.24(s, 3H) 366, 368[M + H]⁺ 70

1H-NMR(400 MHz, DMSO-d6): δ 9.73(s, 1H), 8.50-8.46(m, 1H), 8.30(s, 3H),7.78 (s, 1H), 7.41-7.39(m, 1H), 7.27-7.21(m, 2H), 4.45-4.29(m, 2H),4.06(m, 1H), 2.69 (s, 3H) 318[M + H]⁺

TABLE 11 Example MS Number Structure 1H-NMR (ESI, m/z) 71

1H-NMR(400 MHz, DMSO-d6): δ 8.53(s, 1H), 8.43(s, 1H), 8.28(8,3H),7.81(s, 1H), 7.13(s, 1H), 4.43-4.34(m, 2H), 4.13 (m, 1H), 2.69(s,3H) 368, 370[M + H]⁺ 72

368[M + H]⁺ 73

1H-NMR(300 MHz, DMSO-d6): δ 10.5(s, 1H), 8.38(s, 3H), 7.88-7.86(m, 3H),7.69- 7.49(m, 5H), 7.34-7.29(m, 2H), 4.44-4.43 (m, 2H), 4.22(m, 1H)444[M + H]⁺ 74

1H-NMR(300 MHz, DMSO-d6): δ 11.1(s, 1H), 10.1(s, 1H), 8.34(s, 3H),7.93-7.89 (m, 3H), 7.69-7.59(m, 3H), 7.21(s, 1H), 4.44-4.40(m, 3H) 494,496[M + H]⁺ 75

1H-NMR(300 MHz, DMSO-d6): δ 10.1(s, 1H), 8.37(s, 3H), 7.91-7.88(m, 2H),7.71- 7.55(m, 4H), 7.46(s, 1H), 7.21(s, 1H), 4.46-4.45(m, 2H),4.33-4.32(m, 1H), 2.13 (s, 3H) 492, 494[M + H]⁺ 76

1H-NMR(300 MHz, DMSO-d6): δ 10.1(s, 1H), 9.41(s, 1H), 7.96(s, 1H),7.07(s, 1H), 4.01-3.98(m, 1H), 2.78(s, 6H), 2.60- 2.58(m, 2H),2.23-2.02(m, 2H) 381, 383[M + H]⁺

TABLE 12 Example MS Number Structure 1H-NMR (ESI, m/z) 77

1H NMR(D2O, 400 MHz): δ: 7.71- 7.64 (m, 1H), 7.48-7.38 (m, 3H), 3.96(dd, J = 6.3, 3.7 Hz, 1H), 3.77 (dd, J = 15.2, 3.8 Hz, 1H), 3.60 (dd, J= 15.2, 6.3 Hz, 1H) 304[M + H⁺] 78

1H NMR(D2O, 400 MHz): δ: 7.71 (d, J = 2.6 Hz, 1H), 7.50 (d, J = 2.6 Hz,1H), 3.79-3.70 (m, 1H), 2.70 (s, 3H), 2.64 (t, J = 7.4 Hz, 2H),2.34-2.09 (m, 2H) 367[M + H⁺] 79

1H NMR(D2O, 400 MHz): δ: 7.67 (d, J = 2.3 Hz, 1H), 7.56 (d, J = 2.3 Hz,1H), 3.83 (dd, J = 15.3, 3.5 Hz, 1H), 3.72 (dd, J = 5.2, 3.5 Hz, 1H),3.57 (dd, J = 15.3, 5.3 Hz, 1H), 2.72 (s, 3H), 2.51 (s, 3H) 366[M + H⁺]80

1H NMR (400 MHz, D2O) δ 7.69 (d, J = 2.33 Hz, 1H), 7.31 (dd, J = 2.32,8.18 Hz, 1H), 7.25 (d, J = 8.27 Hz, 1H), 3.95 (dd, J = 3.7, 6.4 Hz, 1H),3.76 (dd, J = 3.8, 15.2 Hz, 1H), 3.58 (dd, J = 6.4, 15.2 Hz, 1H), 2.47(s, 3H). 318[M + H⁺] 81

1H NMR (400 MHz, D2O) δ 7.60 (d, J = 2.7 Hz, 1H), 7.38 (dd, J = 2.7, 8.9Hz, 1H), 7.07 (d, J = 8.9 Hz, 1H), 3.90 (dd, J = 3.6, 6.5 Hz, 1H), 3.83(s, 3H), 3.77 3.66 (m, 1H), 3.60 3.44 (m, 1H). 334[M + H⁺] 82

1H NMR (400 MHz, D2O) δ 7.99 (d, J = 2.3 Hz, 1H), 7.49 (dd, J = 8.6, 2.3Hz, 1H), 7.07 (d, J = 8.7 Hz, 1H), 3.88-3.83 (m, 4H), 3.77 (dd, J =15.2, 3.5 Hz, 1H), 3.56 (dd, J = 15.3, 6.3 Hz, 1H). 334[M + H⁺] 83

377[M + H⁺]

TABLE 13 Example MS Number Structure 1H-NMR (ESI, m/z) 84

365[M + H⁺] 85

268[M + H⁺] 86

337[M + H⁺] 87

1H NMR(DMSO, 400 MHz): δ: 8.14 (s, 1H), 7.83 (d, J = 8.1 Hz, 1H), 7.39(dd, J = 7.7, 1.1 Hz, 1H), 7.19 (t, J = 7.9 Hz, 1H), 6.91-6.82 (m, 1H),3.85-3.78 (m, 1H), 3.67-3.58 (m, 2H), 2.34 (s, 3H) 282[M + H⁺] 88

302[M + H⁺] 89

332[M + H⁺] 90

1H NMR(DMSO, 400 MHz): δ: 8.28- 8.16 (m, 1H), 7.35 (dd, J = 7.9, 1.2 Hz,1H), 7.31 (d, J = 5.9 Hz, 1H), 6.76 (t, J = 8.0 Hz, 1H), 4.01 (t, J =4.9 Hz, 1H), 3.75-3.63 (m, 1H), 3.54-3.38 (m, 1H) 284[M + H⁺]

TABLE 14 Example MS Number Structure 1H-NMR (ESI, m/z) 91

1H NMR(DMSO, 400 MHz): δ (ppm) 8.72 (s, 1H), 8.27 (s, 1H), 7.54 (d, J =8.5 Hz, 1H), 7.25-7.17 (m, 1H), 7.06 (d, J = 8.6 Hz, 1H), 3.90 (s, 3H),3.85-3.50 (m, 3H) 92

313[M + H⁺] 93

1H NMR (400 MHz, DMSO) δ 2.24 (s, 3H), 3.65-3.40 (m, 3H), 6.91 (s, 1H),7.24 (d, J = 8.3 Hz, 1H), 7.47 (d, J = 7.9 Hz, 1H), 8.12 (s, 1H), 8.46(s, 1H). 282[M + H⁺] 94

1H NMR (400 MHz, DMSO) δ 8.61 (s, 1H), 8.23 (s, 1H), 7.41 (d, J = 8.3Hz, 1H), 7.31-7.09 (m, 1H), 6.85 (d, J = 8.2 Hz, 1H), 3.80-3.72 (m, 1H),3.68-3.55 (m, 1H), 3.49-3.32 (m, 1H). 95

1H NMR (400 MHz, DMSO) δ 3.62- 3.45 (m, 2H), 3.77-3.67 (m, 1H), 6.83 (s,1H), 7.37 (d, J = 8.8 Hz, 1H), 7.53 (dd, J = 8.8, 2.7 Hz, 1H), 7.95 (d,J = 2.7 Hz, 1H), 9.47 (s, 1H). 302[M + H⁺] 96

1H NMR (400 MHz, DMSO) δ: 3.44 (m, 2H), 3.82-3.70 (m, 1H), 6.45- 6.20(m, 1H), 6.73 (d, J = 8.8 Hz, 1H), 7.37 (d, J = 8.6 Hz, 1H), 7.80 (s,1H), 8.61 (br, 1H). 284[M + H⁺] 97

1H NMR (400 MHz, DMSO) δ 3.61- 3.41 (m, 2H), 3.77 (m, 1H), 6.63-6.34 (m,1H), 7.01 (d, J = 9.0 Hz, 1H), 7.47 (dd, J = 9.0, 2.7 Hz, 1H), 7.75 (d,J = 2.7 Hz, 1H), 8.97-8.77 (m, 1H).

TABLE 15 Example MS Number Structure 1H-NMR (ESI, m/z) 98

1H NMR (400 MHz, DMSO) δ 3.63- 3.39 (m, 2H), 3.75(s, 3H), 4.02-3.72 (m,1H), 6.47 (s, 1H), 7.01 (d, J = 9.0 Hz, 1H), 7.47 (dd, J = 9.0, 2.7 Hz,1H), 7.75 (d, J = 2.7 Hz, 1H), 8.86 (s, 1H). 298[M + H⁺] 99

1H NMR (400 MHz, DMSO) δ: 2.42 (s, 3H), 3.55-3.40 (m, 2H), 3.72-3.57 (m,1H), 6.53 (s, 1H), 7.14 (d, J = 8.4 Hz, 1H), 7.46 (dd, J = 8.3, 2.2 Hz,1H), 7.93 (d, J = 2.2 Hz, 1H), 9.03 (s, 1H). 282[M + H⁺] 100

298[M + H⁺] 101

1H NMR (400 MHz, DMSO) δ 3.62- 3.41 (m, 4H), 3.75 (s, 3H), 6.74-6.50 (m,1H), 7.01 (s, 1H), 7.36 (s, 1H), 7.58 (s, 1H), 9.32-9.13 (m, 1H).298[M + H⁺] 102

1H NMR (400 MHz, DMSO) δ 3.65- 3.48 (m, 2H), 3.79 (m, 1H), 6.60-6.45 (m,1H), 7.21-7.15 (m, 1H), 7.62-7.54 (m, 1H), 7.98 (dd, J = 6.5, 2.9 Hz,1H), 9.11 (s, 1H). 286[M + H⁺] 103

1H NMR (400 MHz, DMSO) δ 2.12(s, 3H), 2.41(s, 3H), 3.68-3.44 (m, 2H),3.95-3.77 (m, 1H), 6.61 (br, 1H), 6.89 (s, 1H), 7.88 (s, 1H), 7.96 (s,1H). 332[M + H⁺] 104

1H NMR (400 MHz, DMSO) δ 3.69- 3.45 (m, 2H), 3.90-3.80 (m, 1H), 6.96-6.81 (m, 1H), 7.15-7.03 (m, 1H), 7,22- 7.14 (m, 1H), 8.41 (dd, J = 7.9,2.1 Hz, 1H), 8.55 (s, 1H). 322[M + H⁺]

Test Example I Assessment of CaSR Agonistic Activity

(Preparation of CaSR Gene)

CaSR gene was prepared according to the method described in Example 1 ofWO07/55393. The resulting recombinant plasmid was used to generate humanCaSR-expressing plasmid hCaSR/pcDNA3.1.

(Method for Assessing CaSR Agonist)

293E cells (EBNA1-expressing IIEK293 cells, ATCC No. CRL-10852) werecultured in DMEM (1.0 g/ml Glucose-containing Dulbecco's modified Eaglemedium, Nacalai Tesque) containing 10% bovine fetal serum in thepresence of 250 μg/ml of G418. The cells were seeded on a 10 cm-diameterpetri dish at 1.8×10⁶ cells/15 ml, and left to stand in a CO₂ incubator(5% CO₂, 37° C.) for 24 hours. Thereafter, human CaSR expression plasmidhCaSR/pcDNA3.1 was transfected with transfection reagent Mirus Trans IT293 (Takara Bio). Following static culture in a CO₂ incubator for 24hours, the cells were harvested with 10% bovine fetal serum-containingDMEM and seeded on a poly-D-lysine coat 384 well plate (Falcon) at15,000 cells/well. Following static culture in a CO₂ incubator for 24hours, the medium was removed and the resultant was added with 50μl/well of Ca²⁺ fluorescent indicator Calcium 4 Assay Kit (MolecularDevices) dissolved in an assay buffer (146 mM NaCl, 5 mM KCl, 1 mMMgSO₄, 1 mg/ml Glucose, 20 mM HEPES (PH 7.2), 1.5 mM CaCl₂), and left tostand at 37° C. for an hour and then at room temperature for 30 minutesto allow intake of the indicator. The above-mentioned 384-well plate wastransferred to FLIPR (Molecular Devices) and added with 12.5 μl/well ofa compound dissolved in a 0.1% BSA-containing assay buffer to measure3-minute change in the fluorescence intensity.

(Method for Calculating EC₅₀)

The difference between maximum and minimum fluorescence intensitiesbefore and after compound addition (RFU (Max-Min)) was determined byFLIPR automatic calculation. An activity rate was calculated where RFU(Max-Min) upon addition of a compound at a maximum concentration wasdefined 100% and RFU (Max-Min) upon addition of DMSO as a substitute forthe compound at the same concentration was defined 0%. The rate wassubjected to curve fitting using spreadsheet software XLfit to determineEC₅₀ value, i.e., a compound concentration upon 50% activity rate. Theresults are shown in Tables 16 and 17. From these results, the compoundsof the present invention appear to have good CaSR agonistic activity andare useful as CaSR agonistic agents.

TABLE 16 Example Number EC50 (μM) 4 0.040 6 0.003 12 1.1 14 0.003 150.180 16 0.880 17 0.250 32 1.2 34 0.390 41 0.039 42 13.0 43 7.0 47 0.00458 3.1 60 3.4 62 2.3 67 0.003 76 1.8

TABLE 17 Example Number EC50 (μM) 77 0.018 79 0.313 81 3.7 82 1.0 8321.1 85 1.6 88 2.3 89 1.1 90 1.0 92 1.8 93 5.0 99 15.0 100 5.8 101 3.6104 0.490

Test Example II Effect of Decreasing iPTH in Rat by Single Dose ofIntravenous Administration

(Method) A single dose was given to male SD (IGS) rat underpentobarbital anesthesia from a tail vein to examine the transition ofserum iPTH and serum Ca concentration. Blood was taken before theadministration and 5, 15, 30 and 60 minutes after the administration.

Compound No. 1 (the compound described in Example 6) was dissolved inphysiological saline. Meanwhile, cinacalcet as a control substance wasdissolved in PEG400: Saline=1:1 solution.

The results are shown in FIGS. 1 and 2 . Compound No. 1 in Table 1showed almost the same effect as cinacalcet at 0.1 mg/kg in decreasingserum iPTH and serum Ca. Accordingly, the compound of the presentinvention has an effect of decreasing iPTH, and thus suggested to beuseful as a prophylactic or therapeutic agent for hyperparathyroidism.

Test Example III Effect Against Non-Steroidal Anti-Inflammatory Drug(NSAID)-Induced Small Intestine Inflammation

(Method) To a non-fasting rat, Compound No. 1 or 2 (the compounddescribed in Example 14) (10 mg/kg) or Compound No. 3 (the compounddescribed in Example 48) (3, 10 or 30 mg/kg) was orally administered.After 30 minutes, loxoprofen (60 mg/kg) was orally administered and therat was left for 24 hours. Thirty minutes before the autopsy, 1 ml of 1%(w/w) Evansblue dye was intravenously administered to the rat. The smallintestine (from duodenum to ileum) of the animal euthanized under deepether anesthesia was isolated and immersed in 2% formalin for 10 minutesto fix the small intestine from the serosa side. The small intestine wasdissected from the opposite side of the mesentery to measure the injuryarea (mm²) thereof under a 10× dissecting microscope. T-test orDunnett's test was employed as the statistical test where p<0.05 wasconsidered to indicate significant difference.

The results are shown in FIGS. 3 and 4 . Compound No. 1 significantlyimproved the injury area. In addition, Compounds Nos. 2 and 3 showed atendency to improve the injury area. Therefore, compounds of the presentinvention were shown to be useful as a prophylactic or therapeutic agentfor peptic ulcer.

Test Example IV Effect of CaSR Agonist Against Water Absorption ActionUsing Rat Colon Loop Technique

(Method) Appendix and large intestine were isolated from a male SD (IGS)rat under pentobarbital anesthesia, which were ligated 5 cm below theappendix to prepare a large intestine loop. Immediately after looppreparation, PGE2 (4 μg/ml/kg, SIGMA) was intraperitoneallyadministered. Thirty minutes later, 2 ml of Tyrode's solution (NaCl136.9 mM, KCl 2.7 mM, CaCl₂.2H₂O 1.8 mM, MgCl₂.6H₂O 1.04 mM,NaH₂PO₄.2H₂O 0.04 mM, NaH₂PO₄.2H₂O 0.04 mM, Glucose 5.55 mM, NaHCO₃ 11.9mM) was injected into the prepared loop. An hour later, weight of theloop, weight of the loop after removing the fluid therefrom and the looparea were measured to calculate the weight of liquid remaining in theloop per unit area.

The above-described Compounds Nos. 1-3 and Compound No. 4 (the compounddescribed in Example 47) were used as the test compounds while theagents were dissolved in Tyrode's solution.Remaining liquid measure per unit area (g/cm²)=(weight of loop−weight ofloop after removing fluid therefrom)/loop area.

Water absorption was assessed by calculating water regulating effect (%)according to the following formula.Water regulating effect (%)=100−(remaining liquid measure per unit areaobtained with agent−average remaining liquid measure per base unitarea)/(remaining liquid measure per unit area obtained withvehicle−average remaining liquid measure per base unit area)×100.

The results are shown in FIGS. 5-8 . Compounds Nos. 1, 2, 3 and 4promoted water absorption in a dose-depending manner. Accordingly, thecompound of the present invention were shown to be useful as aprophylactic or therapeutic agent for diarrhea.

INDUSTRIAL APPLICABILITY

A compound of the present invention or a salt thereof, and apharmaceutical agent thereof show a superior CaSR agonistic effect, anduseful as a prophylactic or therapeutic agent for a disease that isameliorated through CaSR activation, in particular, hyperparathyroidism,diarrhea, peptic ulcer or the like. In addition, a compound of thepresent invention or a salt thereof can also be used as seasonings thatimparts kokumi.

This application is a continuation of International Patent ApplicationNo. PCT/JP2011/055033, filed on Mar. 4, 2011, and claims priority toJapanese Patent Application No. 2010-048310, filed on Mar. 4, 2010, bothof which are incorporated herein by reference in their entireties.

The invention claimed is:
 1. A compound of Formula (I) or a saltthereof:

wherein R¹ and R², each independently, represent a hydrogen atom, orunsubstituted C₁₋₆ alkyl; R³ represents a hydrogen atom; R⁴ and R⁵, eachindependently, represent a hydrogen atom, substituted or unsubstitutedC₁₋₆ alkyl; X represents CH₂, an oxygen atom, NH, or a sulfur atom; Yrepresents C═O, SO, SO₂, or C═S; R⁶ represents a hydrogen atom; Grepresents R⁷-substituted phenyl or R⁷-substituted pyridyl, where theR⁷-substituted phenyl or the R⁷-substituted pyridyl may further besubstituted with one or two R⁸; R⁷ represents sulfo or carboxyl; R⁸represents substituted or unsubstituted C₁₋₆ alkyl, halogeno, hydroxy,substituted or unsubstituted C₁₋₆ alkoxy, sulfo, or C₁₋₃alkylcarbonylamino, where they may be different when more than one R⁸exist; Q represents a hydrogen atom, substituted or unsubstituted C₁₋₆alkyl, carboxyl, CONR^(e)R^(f), CONHNHR^(g), COR^(h), phenyl, orsubstituted or unsubstituted oxadiazolyl; R^(e) and R^(f), eachindependently, represent a hydrogen atom, substituted or unsubstitutedC₁₋₆ alkyl, substituted or unsubstituted C₁₋₆ alkylsulfonyl,phenylsulfonyl, substituted or unsubstituted C₃₋₈ cycloalkyl, orhydroxy, or alternatively, R^(e) and R^(f) may integrally formmorpholino; R^(g) represents substituted or unsubstituted C₁₋₆alkylcarbonyl; and R^(h) represents substituted or unsubstituted C₁₋₆alkoxy, provided that when X is methylene or an oxygen atom, Y is C═O,all of R¹-R⁵ are hydrogen atoms, and G is R⁷-substituted phenyloptionally substituted with one or two R⁸, then, Q is a group other thancarboxyl or COR^(h).
 2. The compound according to claim 1, or a saltthereof, wherein R⁸ represents substituted or unsubstituted C₁₋₆ alkyl,halogeno, hydroxy, substituted or unsubstituted C₁₋₆ alkoxy, nitro, orsulfo, Q represents a hydrogen atom, substituted or unsubstituted C₁₋₆alkyl (wherein the substituent on said substituted C₁₋₆ alkyl isselected from the group consisting of phenyl, carbamoyl, and cyano),carboxyl, CONR^(e)R^(f), CONHNHR^(g), COR^(h), phenyl, or substituted orunsubstituted oxadiazolyl (wherein the substituent on said substitutedoxadiazolyl is selected from the group consisting of C₁₋₆ alkyl, C₁₋₆alkyl substituted with phenyl, phenyl substituted with halogen, phenylsubstituted with C₁₋₆ alkyl, and thiazolyl); R^(e) and R^(f), eachindependently, represent a hydrogen atom, substituted or unsubstitutedC₁₋₆ alkyl (wherein the substituent on said substituted C₁₋₆ alkyl isselected from the group consisting of phenyl, phenyl substituted withC₁₋₆ alkyl, phenyl substituted with halogen, and imidazolyl), C₁₋₆alkylsulfonyl, phenylsulfonyl, substituted or unsubstituted C₃₋₈cycloalkyl, or hydroxy, or alternatively, R^(e) and R^(f) may integrallyform morpholino; R^(g) represents substituted or unsubstituted C₁₋₆alkylcarbonyl; and R^(h) represents substituted or unsubstituted C₁₋₆alkoxy, provided that when X is methylene or an oxygen atom, Y is C═O,all of R¹-R⁵ are hydrogen atoms, and G is R⁷-substituted phenyloptionally substituted with one or two R⁸, then, Q is a group other thancarboxyl or COR^(h).
 3. A pharmaceutical composition, comprising: thecompound or a pharmaceutically acceptable salt thereof according toclaim 1 or 2 as an active ingredient; and a pharmaceutically acceptableexcipient.
 4. The compound according to claim 1, wherein G representsR⁷-substituted phenyl or R⁷-substituted pyridyl, where theR⁷-substituted phenyl or the R⁷-substituted pyridyl is not furthersubstituted with one or two R⁸.
 5. The pharmaceutical compositionaccording to claim 3, wherein G represents R⁷-substituted phenyl orR⁷-substituted pyridyl, where the R⁷-substituted phenyl or theR⁷-substituted pyridyl is not further substituted with one or two R⁸. 6.The compound according to claim 1, wherein X represents NH, Y representsC═O or C═S, G represents R⁷-substituted phenyl, which may further besubstituted with one or two R⁸, R⁷ represents sulfo or carboxyl, R⁸represents C₁₋₆ alkyl, halogen, hydroxy, C₁₋₃ alkylcarbonylamino orsulfo, where they may be different when more than one R⁸ exist, and Qrepresents carboxyl, CONR^(e)R^(f), CONHNHR^(g) or COR^(h).
 7. Thecompound according to claim 1, wherein G is phenyl, X is NH, Y is CO,and Q is COOH.
 8. The compound according to claim 2, wherein Xrepresents NH, Y represents C═O or C═S, G represents R⁷-substitutedphenyl, which may further be substituted with one or two R⁸, R⁷represents sulfo or carboxyl, R⁸ represents C₁₋₆ alkyl, halogeno,hydroxy, C₁₋₃ alkylcarbonylamino or sulfo, where they may be differentwhen more than one R⁸ exist, and Q represents carboxyl, CONR^(e)R^(f),CONHNHR^(g) or COR^(h).
 9. The compound according to claim 2, wherein Gis phenyl, X is NH, Y is CO, and Q is COOH.
 10. The compound accordingto claim 1, wherein R¹ and R² represent a hydrogen atom, R⁴ and R⁵represent a hydrogen atom, X represents NH, Y represents C═O, Grepresents R⁷-substituted phenyl, which may further be substituted withone or two R⁸, R⁷ represents sulfo, and Q represents COOH.
 11. Thecompound according to claim 1, wherein R¹ and R² represent a hydrogenatom, R⁴ and R⁵ represent a hydrogen atom, X represents NH, Y representsC═O, G represents R⁷-substituted phenyl, which is substituted with twoR⁸, R⁷ represents sulfo, and Q represents COOH.
 12. The compoundaccording to claim 1, wherein R¹ and R² represent a hydrogen atom, R⁴and R⁵ represent a hydrogen atom, X represents NH, Y represents C═O, Grepresents R⁷-substituted phenyl, which is not substituted with R⁸, R⁷represents sulfo, and Q represents COOH.
 13. The compound represented bythe following formula:

or a salt thereof.
 14. A sodium salt of the compound according to claim13.
 15. The compound represented by the following formula:

or a salt thereof.
 16. A sodium salt of the compound according to claim15.
 17. The compound represented by the following formula:

or a salt thereof.
 18. A sodium salt of the compound according to claim17.
 19. The compound represented by the following formula:

or a salt thereof.
 20. A sodium salt of the compound according to claim19.